PheroFIP 19 Joint Meeting of the IOBC/WPRS Working Groups

“Pheromones and other semiochemicals in integrated production” & “Integrated Protection of Fruit Crops”

20-25 January 2019

Lisbon, Portugal

Book of Abstracts

General Index

Welcome to PheroFIP 2019 3 Committees 4 Sponsors 5 Schedule 6 Program 7 Posters presentations 13 Oral Presentations Abstracts index 17 Posters presentations abstracts index 22 Oral Presentations Abstracts 26 Session 1 - Managing beneficial insects by semiochemicals 27 Session 2 - New developments in pest monitoring methods 34 Session 3 - Allelochemical-based pest management tactics 42 Session 4 - Pheromone-based pest management tactics 56 Session 5 - SIT and Biological control of fruit crop pests 73 Session 6 - Applied Biotremology: a new discipline for pest management Vibrational signals as semiophysicals 85 Session 7 - Invasive pests: a challenge for IPM A - Brown Marmorated Stink Bug and other invasive hemipterans 93 B - Spotted Wing Drosophila and other invasive dipterans 104 Session 8 - Integrated protection of fruit crops 125 Posters Presentations Abstracts 136 Session1 - Managing beneficial insects by semiochemicals 137 Session 2 - New developments in pest monitoring methods 139 Session 3 - Allelochemical-based pest management tactics 156 Session 4 - Pheromone-based pest management tactics 173 Session 5 - SIT and Biological control of fruit crop pests 188 Session 6 - Applied Biotremology: a new discipline for pest management. Vibrational signals as semiophysicals 197 Session 7 - Invasive pests: a challenge for IPM A - Brown Marmorated Stink Bug and other invasive hemipterans 200 B - Spotted Wing Drosophila and other invasive dipterans 210 Session 8 - Integrated protection of fruit crops 214

PheroFip 2019 2

WELCOME TO PHEROFIP 2019

Dear Colleagues,

It is with great pleasure that we welcome you to the beautiful city of Lisbon for the Joint Meeting of the IOBC/WPRS Working Groups: Pheromones and other semiochemicals in IP & Integrated Protection of Fruit Crops, PheroFIP 2019.

First, we would like to thank all of you for your participation at the Meeting. We are aware that many of you have travelled a long way to reach us. Thank you!

In this Joint Meeting we are proud to have an outstanding scientific program, structured in 8 sessions, reporting the main developments in semiochemicals and in Integrated Protection of Fruit Crops. In total we have 62 oral presentations (including a plenary talk and 7 Key-note talk presentations) with recent relevant advances. In addition, the meeting will host Flash talks for poster presentations, organized by each session, where the attendees will be able to present their recent research.

This, together with a very nice social program with an excursion to Alentejo region (including a tour to the cellars and wineries and a visit to the UNESCO World Heritage city of Évora) and a Social Dinner at the emblematic Casa do Alentejo, and the well- known attractions of the city of Lisbon, will certainly make your participation at the conference an unforgettable one.

All this has been possible thanks to your participation, to our sponsors support and to the involvement of many volunteer helpers.

We do hope that you enjoy your attendance at the PheroFIP 2019!

José Carlos Franco and Manuela Branco - Conference chairs Jürgen Gross – Convenor Claudio Ioriatti – Convenor Petros Damos – subgroup Convenor Tim Beliën– subgroup Convenor

PheroFip 2019 3

COMMITTEES

SCIENTIFIC COMMITTEE

Ally Harari (Agricultural Research Organization, Israel) Andrea Lucchi (University of Pisa, Italy) António Mexia (University of Lisbon, Portugal) Claudio Ioriatti (Center for Technology Transfer, Fondazione Edmund Mach, Italy) Greg Krawczyk (Pennsylvania State University, USA) Herman Helsen (Wageningen Plant Research, The Netherlands) Howard Thistlewood (Summerland Research and Development Centre, Agriculture and Agri- Food Canada) José Carlos Franco (University of Lisbon, Portugal) Jürgen Gross (Julius Kühn-Institut, Federal Research Centre for Cultivated Plants, Germany) Kerstin Krüger (University of Pretoria, South Africa) Lucía Escudero-Colomar (IRTA, Spain) Manuela Branco (University of Lisbon, Portugal) Marco Tasin (Swedish University of Agricultural Science) Maria Rosa Paiva (Universidade Nova de Lisboa, Portugal) Michele Fountain (NIAB East Malling Research, UK) Nikolaos T. Papadopoulos (University of Thessaly, Greece) Petros Damos (Aristotle University of Thessaloniki, Greece) Thomas Thomidis (Alexander TEI of Thessaloniki, Greece) Tim Beliën (Department of Zoology pcfruit vzw, Belgium)

ORGANISING COMMITTEE

IOBC Coordination Claudio Ioriatti (Center for Technology Transfer, Fondazione Edmund Mach, Italy) Jürgen Gross (Julius Kühn-Institut, Federal Research Centre for Cultivated Plants, Germany)

Local Organization Ana Paula Ramos (LEAF-ISA, University of Lisbon, Portugal) André Garcia (CEF-ISA, University of Lisbon, Portugal) Catarina Mourato (ISA-University of Lisbon, Portugal) Elisabete Figueiredo (LEAF-ISA, University of Lisbon, Portugal) Elsa Borges da Silva (CEF-ISA, University of Lisbon, Portugal) Gonçalo Duarte (LEAF-ISA, University of Lisbon, Portugal) José Carlos Franco (CEF-ISA, University of Lisbon, Portugal) Manuela Branco (CEF-ISA, University of Lisbon, Portugal) Vera Zina (CEF-ISA, University of Lisbon, Portugal)

PheroFip 2019 4

SPONSORS

SUPPORTED BY

PheroFip 2019 5

SCHEDULE

January January January January January January 20 21 22 23 24 25 Session 4 Session 7A Session 8 08:30-08:45 S4.1 S7A.1 S8.1 08:45-09:00 Key-note talk Key-note talk S8.2 Opening 09:00-09:15 S4.2 S7A.2 S8.3 Ceremony 09:15-09:30 Key-note talk S7A.3 S8.4 09:30-09:45 S4.3 S7A.4 S8.5 09:45-10:00 S4.4 S7A.5 S8.6 10:00-10:15 S4.5 S7A.6 Coffee break Coffee break 10:15-10:30 Discussion Discussion 10:30-10:45 S8.7 Coffee break Coffee break 10:45-11:00 Session 1 S8.8 S1.1 Session 7B 11:00-11:15 S4.6 Plenary talk S7B.1 Discussion 11:15-11:30 S4.7 S7B.2 11:30-11:45 S1.2 S4.8 S7B.3 Flash talks* 11:45-12:00 S1.3 S4.9 S7B.4 12:00-12:15 S1.4 S4.10 S7B.5

12:15-12:30 Discussion Discussion Discussion Lunch 12:30-13:30 Lunch Lunch 13:30-14:00 Session 2 Session 5 14:00-14:15 S7B.6 S2.1 S5.1 Business 14:15-14:30 S2.2 Key-note talk S7B.7 Session 14:30-14:45 S2.3 S5.2 S7B.8

14:45-15:00 S2.4 Key-note talk S7B.9 Excursion toAlentejo Excursion 15:00-15:15 S2.5 S5.3 S7B.10 Closing 15:15-15:30 Discussion S5.4 S7B.11 Ceremony 15:30-15:45 S5.5 S7B.12 Awards & Coffee break 15:45-16:00 S5.6 S7B.13 Conclusions 16:00-16:15 Session 3 Discussion S3.1 Discussion 16:15-16:30 Key-note talk Coffee break 16:30-16:45 S3.2 Coffee break 16:45-17:00 S3.3 Session 6 S3.4 S6.1 17:00-17:15 Flash talks* Key-note talk

17:15-17:30 S3.5 S6.2 17:30-17:45 S3.6 S6.3 17:45-18:00 S3.7 Discussion

18:00-18:15 Cocktail Discussion Flash talks* Registration, Registration, 18:15-18:30 and reception Flash talks* 18:30-19:30 19:30-20:00 20:00 Social dinner

*Poster presentations

PheroFip 2019 6

PROGRAM

Sunday, 20th January 17:00 – 19:00 Registration and Reception 18:30 – 19:30 Welcome cocktail

Monday, 21th January 08:30 – 10:00 Opening Ceremony 10:00 – 10:30 Coffee break

SESSION 1 - MANAGING BENEFICIAL INSECTS BY SEMIOCHEMICALS CHAIRPERSON: ANDREA LUCCHI 10:30 – 11:30 PLENARY TALK: S1.1 Protection of Crops against Insect Pests using Chemical Ecology for Managing Beneficial Insects. John Pickett 11:30 – 11:45 S1.2 Development of a biodegradable formulation to enhance aphid biological control. Gunda Thöming, Joakim Pålsson, Teun Dekker, Marco Tasin 11:45 – 12:00 S1.3 Using plant volatiles to monitor natural enemies in horticultural crops. Peter W. Shearer, Vincent P. Jones, Nicholas J. Mills, Kaushalya Amarasekare, Claudio Ioriatti, Andrea Lucchi 12:00 – 12:15 S1.4 Effect of olfactory conditioning on host-searching behaviour of biological control agents. Olga Kostenko, H. Marjolein Kruidhof, Hans M. Smid, Louise E.M. Vet 12:15 – 13:30 DISCUSSION 12:30 – 14:00 LUNCH

SESSION 2 - NEW DEVELOPMENTS IN PEST MONITORING METHODS CHAIRPERSON: TIM BELIEN 14:00 – 14:15 S2.1 An insect olfactomics platform to support rational design of new lures – tephritids as test case. Teun Dekker, Tibebe Dejene, Sebastian Larsson Herrera 14:15 – 14:30 S2.2 Development of multicomponent bisexual kairomone lures to improve management of lepidopteran pests impacting the Canadian codling moth sterile insect program. Gary J.R. Judd, Alan Knight 14:30 – 14:45 S2.3 Evaluating the trap efficiency of two pheromone compounds of male bean flower thrips in the field in Kenya. Falko Drijfhout, David K. Mfuti, Adeyemi O. Akinyemi, Amanuel Tamiru, Sevgan Subramanian, Tom W. Pope, Heather Campbell, William D. J. Kirk 14:45 – 15:00 S2.4 Improving Grapholita molesta monitoring in peach and nectarine orchards under Mating Disruption by using bisexual lures. Michele Preti, Alan L. Knight, Sergio Angeli 15:00 – 15:15 S2.5 Herbivory-induced plant volatiles from apple attract Archips xylosteana (Lepidoptera: Tortricidae). Zaid Badra, Marco Tasin, Sergio Angeli 15:15 – 15:30 DISCUSSION 15:30 – 16:00 COFFEE BREAK & POSTERS

PheroFip 2019 7

SESSION 3 - ALLELOCHEMICAL-BASED PEST MANAGEMENT TACTICS CHAIRPERSON: MICHELLE FOUNTAIN 16:00 – 16:30 KEY-NOTE TALK S3.1 Allelochemicals, how useful are they in plant protection? Olle Anderbrant 16:30 – 16:45 S3.2 Interfering host location of Cacopsylla pruni with repellent plant volatiles, Jannicke Gallinger, Cornelia Dippel, Jürgen Gross 16:45 – 17:00 S3.3 Host defense chemicals as a repellent to Xylosandrus germanus ambrosia beetle infestations in apple trees. Arthur Agnello, Kenneth Lamm 17:00 – 17:15 S3.4 Attractiveness of different lure and infect devices for thrips control. Robert W.H.M. van Tol, Melanie M. Davidson, Ruth C. Butler, David A.J. Teulon, Willem Jan de Kogel 17:15 – 17:30 S3.5 Push-Pull with Synthetic Semiochemicals for Control of Fruit Pests. Michelle T Fountain, Chantelle Jay, Adam Walker, Francesco Rogai, Dudley Farman, David Hall 17:30 – 17:45 S3.6 Sucked in! Crop borders to reduce aphid-transmitted viruses. Kerstin Krüger, Michelle L. Schröder, Robert Glinwood, Rickard Ignell 17:45 – 18:00 S3.7 A new bisexual kairomone lure for codling moth. Alan Knight, Valentina Mujica, Marco Tasin 18:00 – 18:15 DISCUSSION 18:15 – 18:30 FLASH TALKS (POSTER PRESENTATIONS)

Tuesday, 22th January

SESSION 4 - PHEROMONE-BASED PEST MANAGEMENT TACTICS CHAIRPERSON: ALLY HARARI 08:30 – 09:00 KEY-NOTE TALK S4.1 History of a Mating Disruption Mega-dispenser: MSTRS Aerosols and Baggies. Tom Baker 09:00 – 09:30 S4.2 Insect pheromones in Portugal for crop protection: history and present situation. Carlos Frescata 09:30 – 09:45 S4.3 Studies on Lobesia botrana disruption mechanisms based on response after pheromone exposition. Aitor Gavara, Sandra Vacas, Jaime Primo, Vicente Navarro-Llopis 09:45 – 10:00 S4.4 Management of the honeydew moth by mating disruption in vineyard. Patricia Acín 10:00 – 10:15 S4.5 Field effectiveness of pheromone-based mating disruption to control the vine mealybug, Planococcus ficus (Hemiptera: Pseudococcidae): results from Italy. Andrea Lucchi, Pompeo Suma, Edith Ladurner 10:15 – 10:30 DISCUSSION 10:30 – 11:00 COFFEE BREAK & POSTERS

11:00 – 11:15 S4.6 Incorporating Mating Disruption into IPM Programs for Navel Orangeworm in California Almonds. David Haviland, Jhalendra Rijal 11:15 – 11:30 S4.7 On the shoulders of giants: mating disruption of Cydia latiferreana in hazelnuts. Betsey Miller, Daniel Dalton, Vaughn Walton

PheroFip 2019 8

11:30 – 11:45 S4.8 Trials on Pheromone Mating disruption for reducing population of the Western Corn Rootworm, Diabrotica virgifera virgifera, (Coleoptera: Chrysomelidae). Magda Rak Cizej, Silvo Žveplan, Iris Škerbot 11:45 – 12:00 S4.9 Predicting pine processionary moth outbreaks with pheromone trapping. H. Jactel, I. van Halder, E. Kersaudy, T. Belouard 12:00 – 12:15 S4.10 The eucalyptus weevil Gonipterus platensis (Coleoptera, Curculionidae): new control perspectives based on semiochemicals. Sofia Branco, Eduardo Mateus, Stefan Schütz, Maria Rosa Paiva 12:15 – 12:30 DISCUSSION 12:30 – 14:00 LUNCH

SESSION 5 - SIT AND BIOLOGICAL CONTROL OF FRUIT CROP PESTS CHAIRPERSON: KERSTIN KRÜGER 14:00 – 14:30 KEY-NOTE TALK S5.1 Area-wide Management of Fruit Flies Using the Sterile Insect Technique. Rui Cardoso Pereira 14:30 – 15:00 KEY-NOTE TALK S5.2 Back to the future: SIT in synergism with biological control. Massimo Cristofaro, Claudio Ioriatti, Gianfranco Anfora, Valerio Mazzoni, Gino Angeli, Silvia Arnone, Gerardo Roselli, Kim Hoelmer, Max Suckling 15:00 – 15:15 S5.3 Optimizing the sterile insect technique for management of codling moth. Larry Gut, Rob Curtiss, Chris Adams 15:15 – 15:30 S5.4 Hoverflies as biological control agents of the rosy apple aphid in Mediterranean areas. Neus Rodríguez-Gasol, Jesús Avilla, Simó Alegre, Georgina Alins 15:30 – 15:45 S5.5 Intraguild predation among natural enemies of Myzus persicae in peach trees in the NE of Spain. Yahana Aparicio, Rosa Gabarra, Judit Arnó 15:45 – 16:00 S5.6 Developing a biological control strategy for Cacopsylla spp. with novel entomopathogenic fungus Pandora sp. Louisa M. Görg, Annette H. Jensen, Jørgen Eilenberg, Jürgen Gross

16:00 – 16:15 DISCUSSION 16:15 – 16:45 COFFEE BREAK & POSTERS

SESSION 6 - APPLIED BIOTREMOLOGY: A NEW DISCIPLINE FOR PEST MANAGEMENT. VIBRATIONAL SIGNALS AS SEMIOPHYSICALS CHAIRPERSON: VALERIO MAZZONI 16:45 – 17:15 KEY-NOTE TALK S6.1 Biotremology: from basic research to application. Meta Virant-Doberlet, Andrej Čokl, Anna Eriksson, Andrea Lucchi, Valerio Mazzoni, Jernej Polajnar 17:15 – 17:30 S6.2 Vibrational Mating Disruption of Scaphoideus titanus: the current state after two years of field experience and benchmark analysis with Pheromonal Mating Disruption. Valerio Mazzoni 17:30 – 17:45 S6.3 Exploiting vibrational communication for more efficient trapping of Halyomorpha halys (Heteroptera: Pentatomidae). Jernej Polajnar, Lara Maistrello, Valerio Mazzoni

PheroFip 2019 9

17:45 – 18:00 DISCUSSION 18:00 – 18:15 FLASH TALKS (POSTER PRESENTATIONS)

Wednesday, 23th January

08:30 – 18:00 EXCURSION TO ALENTEJO

Thursday, 24th January

SESSION 7 - INVASIVE PESTS: A CHALLENGE FOR IPM

A - BROWN MARMORATED STINK BUG AND OTHER INVASIVE HEMIPTERANS CHAIRPERSON: GREG KRAWCZYK 08:30 – 09:00 KEY-NOTE TALK S7A.1 The invasive Halyomorpha halys in Europe: a challenge for integrated fruit production. Lara Maistrello 09:00 – 09:15 S7A.2 Agroecosystem Impacts on Brown Marmorated Stink Bug Pheromone Trap Capture and Damage in North Carolina Apple Orchards. James Walgenbch, Steven Schoof, Javier Gutierrez, David Crowder 09:15 – 09:30 S7A.3 Biocontrol of the invasive Halyomorpha halys. Hillary Peterson, Jared Ali, Greg Krawczyk 09:30 – 09:45 S7A.4 Predatory ability of wild generalist predators on eggs and first instars of Halyomorpha halys. Giacomo Bulgarini, Zaid Badra, Lara Maistrello 09:45 – 10:00 S7A.5 Feeding, Mating, and Flight Dispersal Behavior of the Invasive Pest Species, Lycorma delicatula (Hemiptera: Fulgoridae). T. C. Baker, Andrew J. Myrick 10:00 – 10:15 S7A.6 Alternative methods to manage brown marmorated stink bug, Halyomorpha halys. Greg Krawczyk, Hillary Peterson, Claire Hirt

10:15 – 10:30 DISCUSSION 10:30 – 11:00 COFFEE BREAK & POSTERS

B - SPOTTED WING DROSOPHILA AND OTHER INVASIVE DIPTERANS CHAIRPERSON: HOWARD THISTLEWOOD 11:00 – 11:15 S7B.1 Automatic identification of Drosophila suzukii (Spotted Wing Drosophila, SWD) by a sensor measuring the wingbeat frequency. Tim Belien, Yenisey M. Mendoza, Bart De Ketelaere, Ammar Alhmedi, Frank Mathijs, Wouter Sayes, Rik Clymans, Dany Bylemans 11:15 – 11:30 S7B.2 Comparative study of Drosophila suzukii females’ behavioral responses to fruit odors of two varieties of Vitis vinifera. Amani Alawamleh, Gordana Ðurović, Maria Giovanna Di Stefano, Sonia Ganassi, Massimo Mancini, Felix Wäckers, Gianfranco Anfora, Antonio De Cristofaro

PheroFip 2019 10

11:30 – 11:45 S7B.3 Evidence for a marking pheromone attracting multiple oviposition in Drosophila suzukii. Gianfranco Anfora, Gabriella Tait, Maria Cristina Crava, Daniel Dalton, Silvia Carlin, Vaughn M. Walton, Marco Valerio Rossi Stacconi 11:45 – 12:00 S7B.4 Laboratory studies on potential oviposition deterrents for Drosophila suzukii. Christa Lethmayer, Matthias Wernicke, Sylvia Blümel 12:00 – 12:15 S7B.5 Spatial and temporal distribution of Drosphila suzukii in relation to fruit quality and coexistence with other arthropods, in sweet cherry orchards of British Columbia. Howard M.A. Thistlewood, Amanda Chamberlain, Susanna Acheampong

12:15 – 12:30 DISCUSSION 12:30 – 14:00 LUNCH

CHAIRPERSON: HEIDRUN VOGT 14:00 – 14:15 S7B.6 Demonstration project "Exclusion netting for managing Spotted Wing Drosophila in fruit crops”. Heidrun Vogt, Bianca Boehnke, Kirsten Köppler, Clemens Augel, Alexandra Wichura, Julian Lindstaedt, Jan-Henrik Wiebusch, Adrian Engel, Silke Benz, Jovanka 14:15 – 14:30 S7B.7 Developing a push-pull system to manage Drosophila suzukii. Dong H. Cha, Anna Wallingford, Peter Landolt, Gregory Loeb 14:30 – 14:45 S7B.8 Exploring the potential of mass trapping Drosophila suzukii. Rik Clymans, Vincent Van Kerckvoorde, Luk De Maeyer, Tim Beliën, Patrick De Clercq, Dany Bylemans 14:45 – 15:00 S7B.9 An innovative management approach for spotted wing drosophila (Drosophila suzukii) using an environmentally friendly attract and kill formulation. Urban Spitaler, Flavia Bianchi, Irene Castellan, Guillermo Rehermann, Daniela Eisenstecken, Paul G. Becher, Sergio Angeli, Silvia Schmidt 15:00 – 15:15 S7B.10 Spotted wing drosophila: Extremely meteorosensitive - a base for the development of a Decision Support System. Kirsten Koeppler, Jeannete Jung, Uwe Harzer, Mandy Püffeld, Marion Gradl, Claudia Tebbe, Alicia Winkler, Paolo Raca, Benno Kleinhenz 15:15 – 15:30 S7B.11 Explorative data analysis of a 7-year Drosophila suzukii monitoring program in Southwest Germany. Felix Briem, Anto Raja Dominic, Burkhard Golla, Christoph Hoffmann, Camilla Englert, Annette Herz, Heidrun Vogt 15:30 – 15:45 S7B.12 Three years of monitoring activities for invasive pest fruit flies in Austria. Alois Egartner, Christa Lethmayer, Richard A. Gottsberger, Sylvia Blümel 15:45 – 16:00 S7B.13 Invasive tephritids (Diptera: Tephritidae) threatening European horticultural production and trade. Nikos T. Papadopoulos, Marc De Meyer, David Nestel, Slawomir Lux, Darren J. Kriticos, Stella Papanastasiou, Kostas Zarpas

16:00 – 16:30 DISCUSSION 16:30 – 17:00 COFFEE BREAK & POSTERS 17:00 – 17:15 FLASH TALKS 20:00 SOCIAL DINNER - CASA DO ALENTEJO (DOWNTOWN)

PheroFip 2019 11

Friday, 25th January

SESSION 8 - INTEGRATED PROTECTION OF FRUIT CROPS CHAIRPERSON: PETROS DAMOS 08:30 – 08:45 S8.1 A survey of integrated pest management in apple orchards in Norway. Bjørn Arild Hatteland, Gaute Myren, Iren Lunde Knutsen, Kristin Kvamm-Lichtenfeld, Endre Bjotveit, Nina Trandem 08:45 – 09:00 S8.2 Effect of the apple cultivar on the duration of the development, survival and penetration rate of Grapholita molesta larval stage under field conditions. Carles Amat, Cesca Alcala, Dolors Bosch, Jesús Avilla, César Gemeno, Lucía-Adriana Escudero-Colomar 09:00 – 09:15 S8.3 Long term trials to reduce pesticide use in fruits- results of EUFRUIT project partners. Barbara Egger, Andreas Naef, Christelle Lacroix, Hinrich Holthusen, Jordi Cabrefiga, Franziska Zavagli 09:15 – 09:30 S8.4 Putting IPM in practice in apple: an 8-year Case Study from Southern Sweden. Marco Tasin, Sanja Manduric, Weronika Swiergiel, Joakim Pålsson, Birgitta Rämert, Mario Porcel 09:30 – 09:45 S8.5 Temperature driven stochastic distribution models and simulation of codling moth phenology. Petros Damos, Polyxeni Soulopoulou, Thomas Thomidis 09:45 – 10:00 S8.6 Towards a knowledge-based decision support system for integrated control of woolly apple aphid, Eriosoma lanigerum, with maximal biological suppression by the parasitoid Apheninus mali. Bangels Eva, Alhmedi Ammar, Bylemans Dany, Belien Tim

10:00 – 10:30 COFFEE BREAK & POSTERS

10:30 – 10:45 S8.7 Plum pocket caused by Taphrina pruni can also be problematic in commercial plum growing. Jorunn Børve, Arne Stensvand 10:45 – 11:00 S8.8 Pseudomonas syringae on young sweet cherry trees in Norway. Jorunn Børve, Juliana Irina Spies Perminow, Marcel Wenneker, May Bente Brurberg, Arne Stensvand

11:00 – 11:30 DISCUSSION 11:30 – 12:00 FLASH TALKES (POSTER PRESENTATIONS)

12:00 – 13:30 LUNCH

13:30 – 15:00 BUSINESS SESSION (CONVENOR ELECTION ET AL.) 15:00 – 16:00 CLOSING CEREMONY (AWARDS & CONCLUSIONS)

PheroFip 2019 12

POSTERS PRESENTATIONS

SESSION 1 - MANAGING BENEFICIAL INSECTS BY SEMIOCHEMICALS

S1.P1 Perspectives of multi-species lures for attracting Chrysopidae. Sándor Koczor, Ferenc Szentkirályi, Miklós Tóth

SESSION 2 - NEW DEVELOPMENTS IN PEST MONITORING METHODS

S2.P1 Attractant for the invasive mealybug citrus pest, Delottococcus aberiae (Hemiptera: Pseudococcidae). Sandra Vacas, Ismael Navarro, Javier Marzo, Vicente Navarro-Llopis, Jaime Prim S2.P2 Attracting Lobesia botrana with microbial volatiles: effect of dose and blend ratio. Sebastian Larsson Herrera, Peter Rikk, Gabriella Köblös, Teun Dekker, Bela Peter Molnar, Marco Tasin S2.P3 EAG responses of Lobesia botrana males and females collected from Vitis vinifera and Daphne gnidium to host-plant volatiles and sex pheromone compounds. Alicia Pérez-Aparicio, Luis Miguel Torres-Vila, César Gemeno S2.P4 Observations on flight activity and voltinism of codling moth Cydia pomonella L. in western central part of Latvia in 2016-2018. Edite Jakobsone, Laura Ozolina-Pole S2.P5 Improving the cost efficiency and practice implementation of automated camera-based monitoring of tortricid pests (Cydia pomonella) in pome fruit orchards. Ammar Alhmedi, Tim Belien, Bart De Ketelaere, Dany Bylemans S2.P6 Failure of commercial sex-pheromone monitoring lures in the assessment of codling moth presence: an emerging problem in South Tyrol. Silvia Schmidt, Peter Neulichedl, Josef Österreicher, Paul Pernter, Michele Preti, Sergio Angeli S2.P7 Use of phenylacetonitrile plus acetic acid as a multi-species trapping method to monitor the leafrollers Pandemis spp. and Choristoneura rosaceana along with Cydia pomonella (Lepidoptera: Tortricidae) in apple. Esteban Basoalto, Alan L. Knight, Gary J. R. Judd S2.P8 Improved monitoring of oriental fruit moth (Lepidoptera: Tortricidae) with terpinyl acetate plus acetic acid membrane lures in Uruguayan orchards. María Valentina Mujica, Alan L. Knight S2.P9 The determination of the effectiveness of pheromone traps for the control Box Tree Moth - Cydalima perspectalis in Georgia. Archil Supatashvili, Medea Burjanadze, Giorgi Mamadashvili, Natia Iordanishvili, Beqa Berdzenishvili, Temel Gorkturk S2.P10 Comparison of counting methods for apple leafcurling midge on pheromone traps. Daniel Cormier, Franz Vanoosthuyse, Gérald Chouinard S2.P11 Progress towards identification of a pheromone of the asparagus beetle, Crioceris asparagi (Coleoptera; Chrysomelidae). Steven Harte, Daniel Bray, Sam Brown, Jude Bennison, David Hall

PheroFip 2019 13

SESSION 3 - ALLELOCHEMICAL-BASED PEST MANAGEMENT TACTICS

S3.P1 Investigating the role of leaf color and plant semiochemicals on the behaviour of Cacopsylla pyri. Bruna Czarnobai De Jorge, Hans E. Hummel, Jürgen Gross S3.P2 KLIMAKOM – Insect's communication under climate change. Margit Rid, Christine Becker, Annette Reineke, Jürgen Gross S3.P3 Luring aphid hyperparasitoids in sweet pepper greenhouses. Jetske G. de Boer, Louise E.M. Vet S3.P4 Is globulol mediating attack by male pioneers of Gonipterus platensis (Col., Curculionidae)? Sofia Branco, Eduardo Mateus, Stefan Schütz, Maria Rosa Paiva S3.P5 Attract and Kill for the control of olive fruit fly in Alto Garda Trentino. Massimo Mucci, Mario Baldessari, Franco Michelotti, Serena Giorgia Chiesa, Gino Angeli S3.P6 Morphology and ultrastructure of the antennal sensilla of Sitophilus granarius (Coleoptera: Curculionidae). Suliman A. I. Ali, Mory Mandiana Diakite, Saqib Ali, Man-Qun Wang

SESSION 4 - PHEROMONE-BASED PEST MANAGEMENT TACTICS

S4.P1 Measurement of Lobesia botrana airborne pheromone by high volume air sampling in mating disruption-treated vineyards. Aitor Gavara, Ismael Navarro, Sandra Vacas, Jaime Primo, Vicente Navarro-Llopis S4.P2 Geostatistical Approach for Spatial Distribution Analysis of Lobesia botrana (Den. & Schiff): (Lepidoptera: Tortricidae) in Douro Demarcated Region (DDR). J. Salvação, C. Carlos, A. Ferreira, M. Nóbrega, G. Fonseca, J. Oliveira, D. Gomes, S. Soares, A. Martinho, R. Soares, F. Gonçalves, L. Torres, J. Aranha S4.P3 Development of the dual purpose mating disruption dispenser to control both the European grapevine moth, Lobesia botrana, and the vine mealybug, Planococcus ficus, in vineyards. Akihiro Baba, Erina Ohno, Tatsuya Hojo, Ryuichi Saguchi, Takeshi Kinsho S4.P4 Control of oriental fruit moth, Cydia molesta Busck and peach twig borer Anarsia lineatella Zell. using reduced rate of pheromone dispensers. Hristina Kutinkova, Vasiliy Dzhuvinov, Desislava Stefanova, Radoslav Andreev, Nedyalka Palagacheva, Bill Lingren S4.P5 Control of plum fruit moth, Grafolitha funebrana Tr., by ISOMATE® - OFM TT dispensers in plum orchards of Bulgaria. D. Stefanova, H. Kutinkova, Petar Savov, R. Andreev, N. Palagacheva, M. Tityanov S4.P6 Synthesis of San Jose scale sex pheromone and its application in field mating disruption. Takeshi Kinsho, Akihiro Baba, Yuki Miyake, Yasuhiko Kutsuwada, Tatsuya Fujii S4.P7 First results on mating disruption of the vine mealybug in Portugal. Elsa Borges da Silva, Catarina Mourato, João Vaz, António Mexia, José Carlos Franco S4.P8 The potential of pheromones for controlling Pseudococcus calceolariae (Hemiptera: Pseudococcidae) in fruit crops. Tania Zaviezo, Jan Bergmann, Alda Romero, Ivan Osorio, Fernanda Flores, Carolina Ballesteros

PheroFip 2019 14

SESSION 5 - SIT AND BIOLOGICAL CONTROL OF FRUIT CROP PESTS

S5.P1 The Sterile Insect Technique for Mediterranean fruit fly control: a North-Italy pilot project Serena G. Chiesa, Gino Angeli, Massimo Cristofaro, Silvia Arnone, Claudio Ioriatti S5.P2 Genetic diversity and initial population size affect the early colonization success of Mastrus ridens, a parasitoid used for the control of codling moth. Tania Zaviezo, Carlos Brochero, Sofía Miranda, Thibaut Malausa S5.P3 Parasitization by Aphelinus mali on Eriosoma lanigerum: an IPM and Organic comparison. Serena Giorgia Chiesa, Luca Corradini, Mario Baldessari, Gino Angeli

SESSION 6 - APPLIED BIOTREMOLOGY: A NEW DISCIPLINE FOR PEST MANAGEMENT. VIBRATIONAL SIGNALS AS SEMIOPHYSICALS

S6.P1 Evaluation of the social behavior based on vibrational signals of Philaenus spumarius in semi-field conditions. Imane Akassou, Sabina Avosani, Vincenzo Verrastro, Valerio Mazzoni

SESSION 7 - INVASIVE PESTS: A CHALLENGE FOR IPM A - BROWN MARMORATED STINK BUG AND OTHER INVASIVE HEMIPTERANS

S7A.P1 Establishment and current status of Halyomorpha halys damaging peaches and olives in the prefecture of Imathia in Northern Greece. Petros Damos, Polyxeni Soulopoulou, Thomas Thomidis S7A.P2 Pheromone traps for Halyomorpha halys: a three year comparison in pear orchards. Giacomo Vaccari, Stefano Caruso, Alberto Pozzebon, Lara Maistrello S7A.P3 Managing Halyomorpha halys by means of exclusion netting: trials 2016-2018. Stefano Caruso, Stefano Vergnani, Giacomo Vaccari, Lara Maistrello S7A.P4 First approach to manage the invasive Halyomorpha halys in Italy: a three-year project. Maria Grazia Tommasini, Lara Maistrello, Francesca Masino, Andrea Antonelli, Pier Paolo Bortolotti, Roberta Nannini, Stefano Caruso, Giacomo Vaccari, Luca Casoli, Michele Preti, Marco Montanari, Matteo Landi, Marco Simoni, Stefano Vergnani S7A.P5 Habitat and tree species effects on Trissolcus japonicus (Hymenoptera: Scelionidae) detections in Virginia, USA. Quinn, NF, EJ Talamas, TC Leskey, JC Bergh

B - SPOTTED WING DROSOPHILA AND OTHER INVASIVE DIPTERANS

S7B.P1 Seasonal changes in odour preferences of Spotted Wing Drosophila (SWD) and the impact of feeding and reproductive status. Tim Beliën, Vincent Van Kerckvoorde, Ammar Alhmedi, Rik Clymans, Eva Bangels, Dany Bylemans S7B.P2 Dissecting oviposition ‘decision making’ in Drosophila suzukii: a hierarchical sensory cascade dictates whether and how much to oviposit. Teun Dekker, Paul Becher, Emil Dekker, Joelle Lemmen S7B.P3 Improved ‘Attract & Kill’ control of Drosophila suzukii (spotted wing Drosophila SWD) with feeding enhancers. V. Van Kerckvoorde, R. Clymans,

PheroFip 2019 15

E. Bangels, A. Alhmedi, T. Thys, M. De Ro, J. Bonte, H. Casteels, P. De Clercq, D. Bylemans, T. Belien

SESSION 8 - INTEGRATED PROTECTION OF FRUIT CROPS

S8.P1 First studies of Grapholita molesta insecticide resistance in field populations from Catalonia (NE of Spain). Dolors Bosch, Adriana Escudero, Jesús Avilla S8.P2 Low-residue fruit production in the Lake Constance region – Model orchard systems for integrated crop protection. Diana Zwahlen, Barbara Egger, Esther Bravin S8.P3 Towards improved integrated control of Anthonomus spp. weevils in pome fruit orchards. Eva Bangels, Tim Beliën, Rik Clymans, Eva Bangels, Tom Thys, Dany Bylemans S8.P4 Silver leaf in Norwegian Prunus domestica plum orchards. Jorunn Børve, Venche Talgø, Arne Stensvand

PheroFip 2019 16

ORAL PRESENTATIONS ABSTRACTS INDEX

SESSION 1 - MANAGING BENEFICIAL INSECTS BY SEMIOCHEMICALS………………...27

CHAIRPERSON: ANDREA LUCCHI Protection of Crops against Insect Pests using Chemical Ecology for Managing Beneficial Insects. John Pickett……………………………………………………………...28 Development of a biodegradable formulation to enhance aphid biological control. Gunda Thöming, Joakim Pålsson, Teun Dekker, Marco Tasin..…………………………………………………………………………………………..29 Using plant volatiles to monitor natural enemies in horticultural crops. Peter W. Shearer, Vincent P. Jones, Nicholas J. Mills, Kaushalya Amarasekare, Claudio Ioriatti, Andrea Lucchi…………………………………………………………………………………30 Effect of olfactory conditioning on host-searching behaviour of biological control agents. Olga Kostenko, H. Marjolein Kruidhof, Hans M. Smid, Louise E.M. Vet………………………………………………………………………………………32

SESSION 2 - NEW DEVELOPMENTS IN PEST MONITORING METHODS…………………..34

CHAIRPERSON: TIM BELIEN An insect olfactomics platform to support rational design of new lures – tephritids as test case. Teun Dekker, Tibebe Dejene, Sebastian Larsson Herrera………………………………………………………………………………………....35 Development of multicomponent bisexual kairomone lures to improve management of lepidopteran pests impacting the Canadian codling moth sterile insect program, Gary J.R. Judd, Alan Knight………………………………………………………………………..36 Evaluating the trap efficiency of two pheromone compounds of male bean flower thrips in the field in Kenya. Falko Drijfhout, David K. Mfuti, Adeyemi O. Akinyemi, Amanuel Tamiru, Sevgan Subramanian, Tom W. Pope, Heather Campbell, William D. J. Kirk…………………………………………………………………………………………… 38 Improving Grapholita molesta monitoring in peach and nectarine orchards under Mating Disruption by using bisexual lures. Michele Preti, Alan L. Knight, Sergio Angeli…………………………………………………………………………………………. 39 Herbivory-induced plant volatiles from apple attract Archips xylosteana (Lepidoptera: Tortricidae). Zaid Badra, Marco Tasin, Sergio Angeli……………………………………...41

SESSION 3 - ALLELOCHEMICAL-BASED PEST MANAGEMENT TACTICS……………… .42

CHAIRPERSON: MICHELLE FOUNTAIN Allelochemicals, how useful are they in plant protection? Olle Anderbrant…………………………………………………………………………….43 Interfering host location of Cacopsylla pruni with repellent plant volatiles. Jannicke Gallinger, Cornelia Dippel, Jürgen Gross…………………………………………...45

PheroFip 2019 17

Host defense chemicals as a repellent to Xylosandrus germanus ambrosia beetle infestations in apple trees. Arthur Agnello, Kenneth Lamm…………………………………………………………………………………..48 Attractiveness of different lure and infect devices for thrips control. Robert W.H.M. van Tol, Melanie M. Davidson, Ruth C. Butler, David A.J. Teulon, Willem Jan de Kogel…………………………………………………………………………………...50 Push-Pull with Synthetic Semiochemicals for Control of Fruit Pests. Michelle T Fountain, Chantelle Jay, Adam Walker, Francesco Rogai, Dudley Farman, David Hall……………………………………………………………………………………..51 Sucked in! Crop borders to reduce aphid-transmitted viruses. Kerstin Krüger, Michelle L. Schröder, Robert Glinwood, Rickard Ignell…………………………...52 A new bisexual kairomone lure for codling moth. Alan Knight, Valentina Mujica, Marco Tasin…………………………………………………………………………...54

SESSION 4 - PHEROMONE-BASED PEST MANAGEMENT TACTICS...... 56

CHAIRPERSON: ALLY HARARI History of a Mating Disruption Mega-dispenser: MSTRS Aerosols and Baggies. Tom Baker…………………………………………………………………………………………..57 Insect pheromones in Portugal for crop protection: history and present situation. Carlos Frescata………………………………………………………………………………………..58 Studies on Lobesia botrana disruption mechanisms based on response after pheromone exposition. Aitor Gavara, Sandra Vacas, Jaime Primo, Vicente Navarro- Llopis…………………………………………………………………………………………...60 Management of the honeydew moth by mating disruption in vineyard. Patricia Acín…………………………………………………………………………………………….61 Field effectiveness of pheromone-based mating disruption to control the vine mealybug, Planococcus ficus (Hemiptera: Pseudococcidae): results from Italy. Andrea Lucchi, Pompeo Suma, Edith Ladurner………………………………………………………………63 Incorporating Mating Disruption into IPM Programs for Navel Orangeworm in California Almonds. David Haviland, Jhalendra Rijal…………………………………….64 On the shoulders of giants: mating disruption of Cydia latiferreana in hazelnuts. Betsey Miller, Daniel Dalton, Vaughn Walton………………………………………………………66 Trials on Pheromone Mating disruption for reducing population of the Western Corn Rootworm, Diabrotica virgifera virgifera, (Coleoptera: Chrysomelidae). Magda Rak Cizej, Silvo Žveplan, Iris Škerbot……………………………………………………….68 Predicting pine processionary moth outbreaks with pheromone trapping. H. Jactel, I. van Halder, E. Kersaudy, T. Belouard……………………………………………………………70 The eucalyptus weevil Gonipterus platensis (Coleoptera, Curculionidae): new control perspectives based on semiochemicals. Sofia Branco, Eduardo Mateus, Stefan Schütz, Maria Rosa Paiva……………………………………………………………………………...71

PheroFip 2019 18

SESSION 5 - SIT AND BIOLOGICAL CONTROL OF FRUIT CROP PESTS…………………73

CHAIRPERSON: KERSTIN KRÜGER Area-wide Management of Fruit Flies Using the Sterile Insect Technique. Rui Cardoso Pereira…………………………………………………………………………………………74 Back to the future: SIT in synergism with biological control. Massimo Cristofaro, Claudio Ioriatti, Gianfranco Anfora, Valerio Mazzoni, Gino Angeli, Silvia Arnone, Gerardo Roselli, Kim Hoelmer, Max Suckling ….………………………………………….76 Optimizing the sterile insect technique for management of codling moth. Larry Gut, Rob Curtiss, Chris Adams……………………………………………………………………77 Hoverflies as biological control agents of the rosy apple aphid in Mediterranean areas. Neus Rodríguez-Gasol, Jesús Avilla, Simó Alegre, Georgina Alins…………………………………………………………………………………………….79 Intraguild predation among natural enemies of Myzus persicae in peach trees in the NE of Spain. Yahana Aparicio, Rosa Gabarra, Judit Arnó…………………………………81 Developing a biological control strategy for Cacopsylla spp. with novel entomopathogenic fungus Pandora sp. Louisa M. Görg, Annette H. Jensen, Jørgen Eilenberg, Jürgen Gross………………………………………………………………………83

SESSION 6 - APPLIED BIOTREMOLOGY: A NEW DISCIPLINE FOR PEST MANAGEMENT. VIBRATIONAL SIGNALS AS SEMIOPHYSICALS………………………………………….85

CHAIRPERSON: VALERIO MAZZONI Biotremology: from basic research to application. Meta Virant-Doberlet, Andrej Čokl, Anna Eriksson, Andrea Lucchi, Valerio Mazzoni, Jernej Polajnar …………………...…..86 Vibrational Mating Disruption of Scaphoideus titanus: the current state after two years of field experience and benchmark analysis with Pheromonal Mating Disruption. Valerio Mazzoni………………………………………………………………………….……89 Exploiting vibrational communication for more efficient trapping of Halyomorpha halys (Heteroptera: Pentatomidae). Jernej Polajnar, Lara Maistrello, Valerio Mazzoni...... 91

SESSION 7 - INVASIVE PESTS: A CHALLENGE FOR IPM……………………………….93

A - BROWN MARMORATED STINK BUG AND OTHER INVASIVE HEMIPTERANS CHAIRPERSON: GREG KRAWCZYK The invasive Halyomorpha halys in Europe: a challenge for integrated fruit production. Lara Maistrello……………………………………………………………………….94 Agroecosystem Impacts on Brown Marmorated Stink Bug Pheromone Trap Capture and Damage in North Carolina Apple Orchards. James Walgenbch, Steven Schoof, Javier Gutierrez, David Crowder…………………………………………………………………….96 Biocontrol of the invasive Halyomorpha halys. Hillary Peterson, Jared Ali, Greg Krawczyk……………………………………………………………………………………... 98 Predatory ability of wild generalist predators on eggs and first instars of Halyomorpha halys. Giacomo Bulgarini, Zaid Badra, Lara Maistrello………………………………….100

PheroFip 2019 19

Feeding, Mating, and Flight Dispersal Behavior of the Invasive Pest Species, Lycorma delicatula (Hemiptera: Fulgoridae). T. C. Baker, Andrew J. Myrick……………….…..102 Alternative methods to manage brown marmorated stink bug, Halyomorpha halys. Greg Krawczyk, Hillary Peterson, Claire Hirt…………………………………….……………..103

B - SPOTTED WING DROSOPHILA AND OTHER INVASIVE DIPTERANS……………………... 104 CHAIRPERSON: HOWARD THISTLEWOOD CHAIRPERSON: HEIDRUN VOGT Automatic identification of Drosophila suzukii (Spotted Wing Drosophila, SWD) by a sensor measuring the wingbeat frequency. Tim Belien, Yenisey M. Mendoza, Bart De Ketelaere, Ammar Alhmedi, Frank Mathijs, Wouter Sayes, Rik Clymans, Dany Bylemans…………………………………………………………………………………..… 105 Comparative study of Drosophila suzukii females’ behavioral responses to fruit odors of two varieties of Vitis vinifera. Amani Alawamleh, Gordana Ðurović, Maria Giovanna Di Stefano, Sonia Ganassi, Massimo Mancini, Felix Wäckers, Gianfranco Anfora, Antonio De Cristofaro…………………………………………………………………………….……….106 Evidence for a marking pheromone attracting multiple oviposition in Drosophila suzukii. Gianfranco Anfora, Gabriella Tait, Maria Cristina Crava, Daniel Dalton, Silvia Carlin, Vaughn M. Walton, Marco Valerio Rossi Stacconi…………………………………………………………………………………….….108 Laboratory studies on potential oviposition deterrents for Drosophila suzukii. Christa Lethmayer, Matthias Wernicke, Sylvia Blümel………………………………………….…109 Spatial and temporal distribution of Drosphila suzukii in relation to fruit quality and coexistence with other arthropods, in sweet cherry orchards of British Columbia. Howard M.A. Thistlewood, Amanda Chamberlain, Susanna Acheampong…………………………………………………………………………………..110 Demonstration project "Exclusion netting for managing Spotted Wing Drosophila in fruit crops”. Heidrun Vogt, Bianca Boehnke, Kirsten Köppler, Clemens Augel, Alexandra Wichura, Julian Lindstaedt, Jan-Henrik Wiebusch, Adrian Engel, Silke Benz, Jovanka………………………………………………………………………………………..112 Developing a push-pull system to manage Drosophila suzukii. Dong H. Cha, Anna Wallingford, Peter Landolt, Gregory Loeb………………………………………:..………113 Exploring the potential of mass trapping Drosophila suzukii. Rik Clymans, Vincent Van Kerckvoorde, Luk De Maeyer, Tim Beliën, Patrick De Clercq, Dany Bylemans……………………………………………………………………………………...115 An innovative management approach for spotted wing drosophila (Drosophila suzukii) using an environmentally friendly attract and kill formulation. Urban Spitaler, Flavia Bianchi, Irene Castellan, Guillermo Rehermann, Daniela Eisenstecken, Paul G. Becher, Sergio Angeli, Silvia Schmidt ...……………………………………………………………..116 Spotted wing drosophila: Extremely meteorosensitive - a base for the development of a Decision Support System. Kirsten Koeppler, Jeannete Jung, Uwe Harzer, Mandy Püffeld, Marion Gradl, Claudia Tebbe, Alicia Winkler, Paolo Raca, Benno Kleinhenz……………………………………………………………………………………..118

PheroFip 2019 20

Explorative data analysis of a 7-year Drosophila suzukii monitoring program in Southwest Germany. Felix Briem, Anto Raja Dominic, Burkhard Golla, Christoph Hoffmann, Camilla Englert, Annette Herz, Heidrun Vogt………………………..………120 Three years of monitoring activities for invasive pest fruit flies in Austria. Alois Egartner, Christa Lethmayer, Richard A. Gottsberger, Sylvia Blümel…………………………………………………………………………………………121 Invasive tephritids (Diptera: Tephritidae) threatening European horticultural production and trade. Nikos T. Papadopoulos, Marc De Meyer, David Nestel, Slawomir Lux, Darren J. Kriticos, Stella Papanastasiou, Kostas Zarpas…………………………………………..123

SESSION 8 - INTEGRATED PROTECTION OF FRUIT CROPS…………………………….125

CHAIRPERSON: PETROS DAMOS A survey of integrated pest management in apple orchards in Norway. Bjørn Arild Hatteland, Gaute Myren, Iren Lunde Knutsen, Kristin Kvamm-Lichtenfeld, Endre Bjotveit, Nina Trandem ……………………………………………………………………..126 Effect of the apple cultivar on the duration of the development, survival and penetration rate of Grapholita molesta larval stage under field conditions. Carles Amat, Cesca Alcala, Dolors Bosch, Jesús Avilla, César Gemeno, Lucía-Adriana Escudero- Colomar………………………………………………………………………………………127 Long term trials to reduce pesticide use in fruits- results of EUFRUIT project partners. Barbara Egger, Andreas Naef, Christelle Lacroix, Hinrich Holthusen, Jordi Cabrefiga, Franziska Zavagli…………………………………………………………………………….128 Putting IPM in practice in apple: an 8-year Case Study from Southern Sweden. Marco Tasin, Sanja Manduric, Weronika Swiergiel, Joakim Pålsson, Birgitta Rämert, Mario Porcel………………………………………………………………………………………….130 Temperature driven stochastic distribution models and simulation of codling moth phenology. Petros Damos, Polyxeni Soulopoulou, Thomas Thomidis………………………………………………………………………………………131 Towards a knowledge-based decision support system for integrated control of woolly apple aphid, Eriosoma lanigerum, with maximal biological suppression by the parasitoid Apheninus mali. Bangels Eva, Alhmedi Ammar, Bylemans Dany, Belien Tim…………………………………………………………………………………………….133 Plum pocket caused by Taphrina pruni can also be problematic in commercial plum growing. Jorunn Børve, Arne Stensvand…………………………………………………..134 Pseudomonas syringae on young sweet cherry trees in Norway. Jorunn Børve, Juliana Irina Spies Perminow, Marcel Wenneker, May Bente Brurberg, Arne Stensvand……………………………………………………………………………………...135

PheroFip 2019 21

POSTERS PRESENTATIONS ABSTRACTS INDEX

SESSION 1 - MANAGING BENEFICIAL INSECTS BY SEMIOCHEMICALS……………….137 Perspectives of multi-species lures for attracting Chrysopidae. Sándor Koczor, Ferenc Szentkirályi, Miklós Tóth……………………………………………………………………138

SESSION 2 - NEW DEVELOPMENTS IN PEST MONITORING METHODS…………………139 Attractant for the invasive mealybug citrus pest, Delottococcus aberiae (Hemiptera: Pseudococcidae). Sandra Vacas, Ismael Navarro, Javier Marzo, Vicente Navarro-Llopis, Jaime Primo………………………………………………………………………………….140 Attracting Lobesia botrana with microbial volatiles: effect of dose and blend ratio. Sebastian Larsson Herrera, Peter Rikk, Gabriella Köblös, Teun Dekker, Bela Peter Molnar, Marco Tasin………………………………………………………………………...141 EAG responses of Lobesia botrana males and females collected from Vitis vinifera and Daphne gnidium to host-plant volatiles and sex pheromone compounds. Alicia Pérez- Aparicio, Luis Miguel Torres-Vila, César Gemeno………………………………………..142 Observations on flight activity and voltinism of codling moth Cydia pomonella L. in western central part of Latvia in 2016-2018. Edite Jakobsone, Laura Ozolina-Pole…..143 Improving the cost efficiency and practice implementation of automated camera-based monitoring of tortricid pests (Cydia pomonella) in pome fruit orchards. Ammar Alhmedi, Tim Belien, Bart De Ketelaere, Dany Bylemans………………………………..145 Failure of commercial sex-pheromone monitoring lures in the assessment of codling moth presence: an emerging problem in South Tyrol. Silvia Schmidt, Peter Neulichedl, Josef Österreicher, Paul Pernter, Michele Preti, Sergio Angeli…………………………...146 Use of phenylacetonitrile plus acetic acid as a multi-species trapping method to monitor the leafrollers Pandemis spp. and Choristoneura rosaceana along with Cydia pomonella (Lepidoptera: Tortricidae) in apple. Esteban Basoalto, Alan L. Knight, Gary J. R. Judd………………………………………………………………………………………..….148 Improved monitoring of oriental fruit moth (Lepidoptera: Tortricidae) with terpinyl acetate plus acetic acid membrane lures in Uruguayan orchards. María Valentina Mujica, Alan L. Knight………………………………………………………………………150 The determination of the effectiveness of pheromone traps for the control Box Tree Moth - Cydalima perspectalis in Georgia. Archil Supatashvili, Medea Burjanadze, Giorgi Mamadashvili, Natia Iordanishvili, Beqa Berdzenishvili, Temel Gorkturk………………………………………………………………………………….…..152 Comparison of counting methods for apple leafcurling midge on pheromone traps. Daniel Cormier, Franz Vanoosthuyse, Gérald Chouinard………………………………...153 Progress towards identification of a pheromone of the asparagus beetle, Crioceris asparagi (Coleoptera; Chrysomelidae). Steven Harte, Daniel Bray, Sam Brown, Jude Bennison, David Hall…………………………………………………………………………154

PheroFip 2019 22

SESSION 3 - ALLELOCHEMICAL-BASED PEST MANAGEMENT TACTICS………………156 Investigating the role of leaf color and plant semiochemicals on the behaviour of Cacopsylla pyri. Bruna Czarnobai De Jorge, Hans E. Hummel, Jürgen Gross…………157 KLIMAKOM – Insect's communication under climate change. Margit Rid, Christine Becker, Annette Reineke, Jürgen Gross…………………………………………………….162 Luring aphid hyperparasitoids in sweet pepper greenhouses. Jetske G. de Boer, Louise E.M. Vet……………………………………………………………………………………….166 Is globulol mediating attack by male pioneers of Gonipterus platensis (Col., Curculionidae)? Sofia Branco, Eduardo Mateus, Stefan Schütz, Maria Rosa Paiva……167 Attract and Kill for the control of olive fruit fly in Alto Garda Trentino. Massimo Mucci, Mario Baldessari, Franco Michelotti, Serena Giorgia Chiesa, Gino Angeli……………...170 Morphology and ultrastructure of the antennal sensilla of Sitophilus granarius (Coleoptera: Curculionidae), Suliman A. I. Ali, Mory Mandiana Diakite, Saqib Ali, Man- Qun Wang…………………………………………………………………………………….172

SESSION 4 - PHEROMONE-BASED PEST MANAGEMENT TACTICS……………………..173 Measurement of Lobesia botrana airborne pheromone by high volume air sampling in mating disruption-treated vineyards. Aitor Gavara, Ismael Navarro, Sandra Vacas, Jaime Primo, Vicente Navarro-Llopis………………………………………………………….….174 Geostatistical Approach for Spatial Distribution Analysis of Lobesia botrana (Den. & Schiff): (Lepidoptera: Tortricidae) in Douro Demarcated Region (DDR). J. Salvação, C. Carlos, A. Ferreira, M. Nóbrega, G. Fonseca, J. Oliveira, D. Gomes, S. Soares, A. Martinho, R. Soares, F. Gonçalves, L. Torres, J. Aranha………………………………....175 Development of the dual purpose mating disruption dispenser to control both the European grapevine moth, Lobesia botrana, and the vine mealybug, Planococcus ficus, in vineyards. Akihiro Baba, Erina Ohno, Tatsuya Hojo, Ryuichi Saguchi, Takeshi Kinsho…………………………………………………………………………………………176 Control of oriental fruit moth, Cydia molesta Busck and peach twig borer Anarsia lineatella Zell. using reduced rate of pheromone dispensers. Hristina Kutinkova, Vasiliy Dzhuvinov, Desislava Stefanova, Radoslav Andreev, Nedyalka Palagacheva, Bill Lingren……………………………………………………………………………………….177 Control of plum fruit moth, Grafolitha funebrana Tr., by ISOMATE® - OFM TT dispensers in plum orchards of Bulgaria. D. Stefanova, H. Kutinkova, Petar Savov, R. Andreev, N. Palagacheva, M. Tityanov……………………………………………………..179 Synthesis of San Jose scale sex pheromone and its application in field mating disruption. Takeshi Kinsho, Akihiro Baba, Yuki Miyake, Yasuhiko Kutsuwada, Tatsuya Fujii……………………………………………………………………………………………181 First results on mating disruption of the vine mealybug in Portugal. Elsa Borges da Silva, Catarina Mourato, João Vaz, António Mexia, José Carlos Franco………………………182 The potential of pheromones for controlling Pseudococcus calceolariae (Hemiptera: Pseudococcidae) in fruit crops. Tania Zaviezo, Jan Bergmann, Alda Romero, Ivan Osorio, Fernanda Flores, Carolina Ballesteros….…………………………………………184

PheroFip 2019 23

SESSION 5 - SIT AND BIOLOGICAL CONTROL OF FRUIT CROP PESTS………………..188 The Sterile Insect Technique for Mediterranean fruit fly control: a North-Italy pilot project. Serena G. Chiesa, Gino Angeli, Massimo Cristofaro, Silvia Arnone, Claudio Ioriatti…………………………………………………………………………………………189 Genetic diversity and initial population size affect the early colonization success of Mastrus ridens, a parasitoid used for the control of codling moth. Tania Zaviezo, Carlos Brochero, Sofía Miranda, Thibaut Malausa………………………………………………. 192 Parasitization by Aphelinus mali on Eriosoma lanigerum: an IPM and Organic comparison. Serena Giorgia Chiesa, Luca Corradini, Mario Baldessari, Gino Angeli…194

SESSION 6 - APPLIED BIOTREMOLOGY: A NEW DISCIPLINE FOR PEST MANAGEMENT VIBRATIONAL SIGNALS AS SEMIOPHYSICALS………………………………………...197 Evaluation of the social behavior based on vibrational signals of Philaenus spumarius in semi-field conditions- Imane Akassou, Sabina Avosani, Vincenzo Verrastro, Valerio Mazzoni……………………………………………………………………………………….198

SESSION 7 - INVASIVE PESTS: A CHALLENGE FOR IPM………………………………200

A - BROWN MARMORATED STINK BUG AND OTHER INVASIVE HEMIPTERANS Establishment and current status of Halyomorpha halys damaging peaches and olives in the prefecture of Imathia in Northern Greece. Petros Damos, Polyxeni Soulopoulou, Thomas Thomidis…………………………………………………………………………….201 Pheromone traps for Halyomorpha halys: a three year comparison in pear orchards. Giacomo Vaccari, Stefano Caruso, Alberto Pozzebon, Lara Maistrello………………….203 Managing Halyomorpha halys by means of exclusion netting: trials 2016-2018. Stefano Caruso, Stefano Vergnani, Giacomo Vaccari, Lara Maistrello…………………………...205 First approach to manage the invasive Halyomorpha halys in Italy: a three-year project. Maria Grazia Tommasini, Lara Maistrello, Francesca Masino, Andrea Antonelli, Pier Paolo Bortolotti, Roberta Nannini, Stefano Caruso, Giacomo Vaccari, Luca Casoli, Michele Preti, Marco Montanari, Matteo Landi, Marco Simoni, Stefano Vergnani…….207 Habitat and tree species effects on Trissolcus japonicus (Hymenoptera: Scelionidae) detections in Virginia, USA. Quinn, NF, EJ Talamas, TC Leskey, JC Bergh…………..209

B - SPOTTED WING DROSOPHILA AND OTHER INVASIVE DIPTERANS……………………...210 Seasonal changes in odour preferences of Spotted Wing Drosophila (SWD) and the impact of feeding and reproductive status. Tim Beliën, Vincent Van Kerckvoorde, Ammar Alhmedi, Rik Clymans, Eva Bangels, Dany Bylemans………………………..211 Dissecting oviposition ‘decision making’ in Drosophila suzukii: a hierarchical sensory cascade dictates whether and how much to oviposit. Teun Dekker, Paul Becher, Emil Dekker, Joelle Lemmen………………………………………………………………………212 Improved ‘Attract & Kill’ control of Drosophila suzukii (spotted wing Drosophila SWD) with feeding enhancers. V. Van Kerckvoorde, R. Clymans, E. Bangels, A. Alhmedi, T. Thys, M. De Ro, J. Bonte, H. Casteels, P. De Clercq, D. Bylemans, T. Belien………………………………………………………………………………………….213

PheroFip 2019 24

SESSION 8 - INTEGRATED PROTECTION OF FRUIT CROPS…………………………….214 First studies of Grapholita molesta insecticide resistance in field populations from Catalonia (NE of Spain). Dolors Bosch, Adriana Escudero, Jesús Avilla………………..215 Low-residue fruit production in the Lake Constance region – Model orchard systems for integrated crop protection. Diana Zwahlen, Barbara Egger, Esther Bravin…………….216 Towards improved integrated control of Anthonomus spp. weevils in pome fruit orchards. Eva Bangels, Tim Beliën, Rik Clymans, Eva Bangels, Tom Thys, Dany Bylemans………………………………………………………………………………..…….218 Silver leaf in Norwegian Prunus domestica plum orchards. Jorunn Børve, Venche Talgø, Arne Stensvand……………………………………………………………………………….219

PheroFip 2019 25

ORAL PRESENTATIONS ABSTRACTS

PheroFip 2019 26

SESSION 1

MANAGING BENEFICIAL INSECTS BY

SEMIOCHEMICALS

CHAIRPERSON: ANDREA LUCCHI

PheroFip 2019 27

Protection of Crops against Insect Pests using Chemical Ecology for Managing Beneficial Insects

John A Pickett1

1School of Chemistry, Cardiff University, Wales, UK

Abstract From an understanding of the chemical ecology of tritrophic interactions we can exploit natural populations of entomophagous insects, such as hymenopterous parasitoids, for sustainable pest management. There are two prerequisites for such sustainability, which are that the natural parasitoid populations need to be maintained and that foraging by the parasitoids needs to be enhanced before pest threshold populations are reached. Success in this approach is evidenced in the push-pull system developed for sub-Saharan cereal crop protection against lepidopterous stemborers. Natural parasitoid populations are maintained by botanically diverse agro-ecological conditions incorporating wild host plants of the herbivores. The parasitoids are then lured into the cereal crops by intercrops releasing foraging semiochemicals, such as the plant stress related C11 and C16 homoterpenes naturally induced by feeding on the crop plant hosts. For industrialised arable agriculture, induction or priming mechanisms are targeted and these processes will need to be managed, including via GM, by use of plant defence elicitors applied to the crop or released by sentinel plants. However, for high value horticultural crops, such as for fruit production, agro-ecological approaches can be further developed. The potential depletion of natural parasitoid populations needs to be accommodated by increased ecosystem services in the agro-ecological regions involved and will be costed as part of a “land sparing” rather than “land sharing” policy. In addition to the use of foraging cues from crop plants, signals for dispersing parasitoids from production sites created by land sparing are required and may be obtained from signalling involving systems at the fourth trophic level. Certain multitrophic signalling systems involving highly unstable semiochemicals, including certain sesquiterpenes, are currently being developed further by synthetic biological approaches.

PheroFip 2019 28

Development of a biodegradable formulation to enhance aphid biological control

Gunda Thöming1, Joakim Pålsson2, Teun Dekker2, Marco Tasin2

1NIBIO, Norwegian Institute of Bioeconomy Research, 1431 Ås, Norway; 2SLU, Department of Plant Protection Biology, 230 53 Alnarp, Sweden.

Abstract Although supporting high productivity, modern agriculture caused a long-term impact on natural trophic interactions, releasing pests from pressure linked with their natural enemies. Studies have demonstrated that volatiles released under herbivory can recruit natural enemies of pests from a distance. Here, we used a novel biodegradable formulation loaded with induced and food-signalling volatiles with the aim to attract the green lacewing, Chrysoperla carnea, and increase biological control of two cereal aphids Sitobion avenae and Rhopalosiphum padi. The new product consisted of a biodegradable matrix loaded with a 3- component blend of methyl salicylate, acetic acid and phenylacetaldehyde in a 1:1:1 ratio. Field experiments were carried out in a barley field in Norway. Single plants were provided with a 1 ml dollop of the new formulation or with a standard polyethylene emballage dispenser loaded with the same amount of compounds. The number of lacewing eggs and larvae as well as the attraction of additional natural enemies was recorded both on the treated and surrounding plants by visual inspection. At the same time, an assessment of aphid infestation was carried out. A higher local density of lacewing adults, eggs and larvae over an 8-week period was observed for both the standard and the biodegradable formulation in comparison with untreated plants. Chemical analysis of the volatiles emitted from the slow- release matrix showed an active emission of the blend over at least a 4-week period. Significant biological control of aphid was measured in the vegetation surrounding the odour source. Both aphid populations were significantly reduced, with no difference between the new and the standard treatment. While coccinellids and hoverflies were not affected by the treatment, a lower number of mummified aphids were measured in some of the treated plants in comparison with untreated ones. Results show the potential for semiochemical-based targeted attraction of lacewings to enhance biological control of aphids in a prevalent monoculture field setting. Additional studies are required to support the development of practical integrated pest management guidelines, including optimization of application density, threshold value for pest and natural enemies and practical recommendation for the establishment of non-crop vegetation within and around the crop.

Key words: Chrysoperla carnea, integrated pest management, kairomone, barley

PheroFip 2019 29

Using plant volatiles to monitor natural enemies in horticultural crops

Peter W. Shearer1, Vincent P. Jones2, Nicholas J. Mills3, Kaushalya Amarasekare4, Claudio Ioriatti5, Andrea Lucchi6

1Strawberry Center, California Polytechnic State University, San Luis Obispo, CA 93407, USA 2Department of Entomology, Washington State University Tree Fruit Research and Extension Center, Wenatchee, WA 98801, USA; 3Department of Environmental Science and Policy Management, University of California, Berkeley, CA 94720-3114, USA; 4Department of Agricultural and Environmental Sciences, Tennessee State University, Nashville, TN, 37209, USA; 5Centre for Technology Transfer, Fondazione Edmund Mach, San Michele a/A, Trento, Italy; 6Department Agriculture, Food and Environment, University of Pisa, Italy

Abstract In November, 2016, the Journal Biological Control released a Special Issue, “Enhancing biological control in western orchards: a five-year project”, which was devoted entirely to reporting results from a United States Department of Agriculture, National Institute of Food and Agriculture (USDA-NIFA) Specialty Crop Research Initiative (SCRI) grant (Biological Control 2016). The effort was designed to help improve biological control in apple, pear and walnut orchards in the western USA that were transitioning from older insecticides towards organophosphorus (OP) replacement and reduced-risk insecticides. One major component of this project included evaluating plant volatiles for monitoring natural enemies in orchards (Jones et al. 2016a). For this study, various combinations of herbivore-induced plant volatiles (HIPV) and floral volatiles were investigated for their attractiveness for monitoring key natural enemies found in orchards in California, Oregon and Washington. Traps baited with plant volatile lures capture flying natural enemies over an extended period of time compared with instantaneous measures that occur with beat trays. Below is a brief synopsis of some of our results. Combinations of attractants that provided the greatest capture varied by natural enemy taxon. Traps baited with lures containing phenylacetaldehyde (PAA) were attractive to Hymenoptera. Syrphids (Diptera: Syrphidae) responded strongly to 2-phenylethanol (PE). The addition of geraniol (GE) increased capture of Eupoides fumipennis while adding acetic acid (AA) to PE decreased capture. Multi-component lures worked best for Chrysoperla spp. (Neuroptera: Chrysopidae). Acetic acid was a common component in traps that captured the most Chrysoperla spp. Capture increased when either PE, PAA or acetophenone (AP) was added to AA. Traps baited with AA+AP+PAA or AA+AP+PE were the optimal combination of attractants for the Chrysoperla complex we studied. Plant volatiles are useful for studying natural enemies and can be used to estimate the diversity of natural enemy communities in various orchard systems (Mills et al. 2016). They can be used to follow changes in natural enemy diversity, richness and evenness among crops and between orchards within crops. They also can be used to develop phenology models for natural enemies (Jones et al. 2016b). Traps baited with plant volatile lures can also be used to measure changes in natural enemy abundance and impacts on natural enemy/prey ratios following applications of insecticides. Shearer et al. 2016 demonstrated that plant volatile baited traps captured more predatory neuroptera in pear plots treated with spinetoram than in plots treated with either chlorantraniliprole or cyantraniliprole. Some of these plant volatiles were tested alongside Malaise traps in Italian vineyards (Lucchi et al. 2017). Traps baited with methyl salicylate, AA and PE were attractive to Chrysopidae, especially to adults in the genus Chrysoperla, but not to Pseudomallada.

PheroFip 2019 30

Insectcapture data from baited traps and Malaise traps revealed that Malaise traps were more efficient at monitoring Syrphidae while baited-sticky traps captured more adult Chrysopidae. In this instance, using different sampling methods improved knowledge of species richness. It is important to note that using traps baited with plant volatiles to monitor natural enemies is more difficult than using sex pheromone baited traps. However, they do have a role for monitoring natural enemy phenology and abundance through time. Additionally, because in our studies these attractants worked across multiple crops over a wide geographical area, they most likely will have utility in other ecosystems.

Key Words: herbivore induced plant volatiles, HIPV, floral volatiles, baited sticky trap, Malaise trap

References Biological Control. 2016. in Jones, V. & Mills, N. (guest eds.): Enhancing biological control in western orchards: a five-year project. Elsevier, Amsterdam, The Netherlands. 102. 127 pp. Jones, V. P., Horton, D. R., Mills, N. J., Unruh, T. R., Miliczky, E., Shearer, P. W., Baker, C. C., & Melton, T. D. 2016a: Evaluating plant volatiles for monitoring natural enemies in apple, pear and walnut orchards. Biological Control 102: 53-65. Jones, V. P., Horton, D. R., Mills, N. J., Unruh, T. R., Miliczky, E., Shearer, P. W., Baker, C. C., Melton, T. D. & Amarasekare, K. 2016b: Using plant volatile traps to develop phenology models for natural enemies: An example using Chrysopa nigricornis (Burmeister) (Neuroptera: Chrysopidae). Biological Control 102: 77-84. Lucchi, A., Loni, A., Gandini, L. M., Scaramozzino, P., Ioriatti, C., Ricciardi, R. & Shearer, P. W. 2017: Using herbivore-induced plant volatiles to attract lacewings, hoverflies and parasitoid wasps in vineyards: achievements and constraints. Bulletin of Insectology 70(2): 273-282. Mills, N. J., Jones, V. P., Baker, C. C., Melton, T. D., Steffan, S. A., Unruh, T. R., Horton, D. R., Shearer, P. W., Amarasekare, K. G. & Milickzy, E. 2016: Using plant volatile traps to estimate the diversity of natural enemy communities in orchard ecosystems. Biological Control 102: 66-76. Shearer, P. W., Amarasekare, K. G., Castagnoli, S. P., Beers, E. H., Jones, V. P. & Mills, N. J. 2016: Large-plot field studies to assess impacts of newer insecticides on non-target arthropods in Western U.S. orchards. Biological Control 102: 26-34.

Acknowledgment The authors acknowledge that the primary work reported here was funded by the United States Department of Agriculture, National Institute of Food and Agriculture (USDA-NIFA) Specialty Crop Research Initiative (SCRI) grant # 2008-04854.

PheroFip 2019 31

Effect of olfactory conditioning on host-searching behaviour of biological control agents

Olga Kostenko1*, H. Marjolein Kruidhof3, Hans M. Smid2, Louise E.M. Vet1,2

1Netherlands Institute of Ecology (NIOO-KNAW), Department of Terrestrial Ecology, PO Box 50, 6700 AB Wageningen, The Netherlands; 2Wageningen UR, Department of Entomology, PO Box 8031, 6700 EH Wageningen, The Netherlands; 3Wageningen UR Greenhouse Horticulture, PO Box 20, 2265 ZG Bleiswijk, The Netherlands *[email protected]

Abstract Over the last decades the use of mass-reared parasitoids has been established as the basis for integrated pest management strategies in many crops. Still, their performance in the greenhouse or field is not always optimal. Inconsistent results in biological pest control may not only result from variation in the foraging environment or the genetic composition of parasitoid populations, but likely depend to a large extent on the naive, inexperienced state of the used parasitoids. When searching for herbivorous host-insects, parasitoids make use of so- called herbivore-induced plant volatiles (HIPVs) that are emitted by the plant in response to insect feeding damage. Upon a first host encounter parasitoids can learn to associate these HIPVs with the presence of the host. As a result of this associative learning, parasitoids may 1) increase or decrease their innate response levels; 2) acquire responses to novel stimuli, 3) shift their preferences towards the learned HIPVs, resulting in temporary specialisation; and/or 4) change movement patterns. Field and laboratory studies have shown that learning improves foraging efficiency of parasitoids (enhances the chance and reduce the time needed to find a host). This holds great promise for the optimization of biological pest control, but it has been rarely explored before. Here we will present the results of several behavioural experiments examining the effect of olfactory conditioning of parasitoids on their searching efficiency for mealybug-infested plants. First, behavioural assays in large cages in an experimental greenhouse demonstrated that overall almost twice as many conditioned Anagyrus pseudococci Girault (Hymenoptera: Encyrtidae) were recaptured on mealybug-infested pot roses compared to unconditioned A. pseudococci parasitoids. A positive effect of olfactory conditioning was manifested already four hours after the release of A. pseudococci parasitoids. While the total number of parasitoids recaptured on mealybug-infested rose plants over a 72-hour period was independent of the plant odour (mealybug-infested rose plant vs mealybug-infested bell- pepper plant) used during olfactory conditioning, the recapture rate of the parasitoids (the number of recaptured parasitoids at different time points) was significantly higher when parasitoids were conditioned on mealybug-infested rose plants. This was particularly the case for the recapture rate of A. pseudococci wasps. Second, several behavioural assays using olfactometers also confirmed that olfactory conditioning increases the response and/or preference of A. pseudococci towards mealybug-infested rose plants. These results suggest that olfactory conditioning can have a favourable effect on the ability of A. pseudococci parasitoids to locate their hosts and therefore can potentially improve the ability of parasitoids to control mealybugs.

Key words: Planococcus citri, roses, parasitoids, associative learning, herbivore-induced plant volatiles, Anagyrus pseudococci

PheroFip 2019 32

Acknowledgements This work was funded by the Netherlands Organization of Scientific research (NWO, “GROEN” project number ALWGR.2015.7) and co-funded by Biobee Biological Systems and Koppert Biological Systems.

PheroFip 2019 33

SESSION 2

NEW DEVELOPMENTS IN PEST

MONITORING METHODS

CHAIRPERSON: TIM BELIEN

PheroFip 2019 34

An insect olfactomics platform to support rational design of new lures – tephritids as test case

Teun Dekker1, Tibebe Dejene1, Sebastian Larsson Herrera1

1Department of Plant Protection Biology, Swedish University of Agricultural Sciences, Sweden

Abstract Decades of research on the sense of smell in insects has not yet produced a public, online database of unified olfactory responses across species. Here we present evidence on how such an olfactome database can be constructed, and used to extract evolutionary-ecological signals, as well as for attractants. We compared four species of tephritid fruit flies, and Drosophila melanogaster as outgroup, in their responses to volatiles from four fruits. We found that sharedness of volatiles is an important explanatory variable in detection, in spite of several hundred of millions years of evolutionary separation. Sharedness also appeared to contain some sort of blueprint of ‘attractiveness’, as a blend of shared volatiles attracted all teprhitid and even the very distantly related D. melanogaster. I will present further expansions on how to utilize this database to contribute to better eco-evo understanding of olfaction as well as its use in understanding ‘attraction’- A draft olfactome database along with ‘on-the-click’ analysis tools is publicly available at www.tephri.org. We welcome collaborations for further developing the olfactome platform.

PheroFip 2019 35

Development of multicomponent bisexual kairomone lures to improve management of lepidopteran pests impacting the Canadian codling moth sterile insect program

Gary Judd1, Alan Knight2*

1Agriculture and Agri-food Canada, Summerland Research and Development Centre, 4200 Highway 97, Summerland, British Columbia, Canada, V0H 1Z0. [email protected]; 2Yakima Agricultural Research Laboratory, USDA-ARS, 5230 Konnowac Pass Rd, Wapato, Washington 98951, USA. *[email protected]

Abstract Codling moth, Cydia pomonella L. (Lepidoptera: Tortricidae) has been the target of an areawide program integrating sterile insect technology (SIT) and pheromone-based mating disruption (MD) in British Columbia, Canada, since 1994. Until very recently, this areawide program had virtually eliminated use of insecticides for codling moth, enabling growth of the largest organic apple industry in Canada, while simultaneously making it vulnerable to a resurgence of native, and an invasion of secondary pests. We are developing lures to improve management of codling moth and several secondary lepidopteran species, by focusing on multicomponent bisexual kairomones. Unlike sex pheromone lures, binary, acetic acid (AA) plus pear ester (PE) kairomone lures attract male and female codling moth and do not compete with the thousands of natural (calling sterile females) and synthetic pheromone sources (MD dispensers) present in this areawide system. Therefore, traps baited with an AA- PE kairomone lure can track densities of sterile moths in relation to the phenology and density of wild moths better, and more effectively measure critical sterile to wild codling moth ratios, than do standard sex pheromone traps. Traps baited with AA-PE kairomone lures also remove fewer functional sterile males than do traps baited with pheromone. AA-PE lures can also be used to monitor male and female moths of the invasive Synanthedon myopaeformis Borkhausen (Lepidoptera: Sesiidae), but their use in developing mass trapping or attract and kill technologies for this pest is somewhat limited because they attract sterile codling moths. By adding a behavioural antagonist to the AA-PE lure, we are able to reduce captures of male codling moth, while retaining attraction of S. myopaeformis. By adding ethyl butyrate (EB), a feeding stimulant for S. myopaeformis, to the AA-PE lure, we can significantly increase catches of S. myopaeformis, without affecting catches of codling moth. When phenylacetonitrile (PAN) or 2-phenylethanol (PET), known tortricid kairomone components (Judd et al. 2017a, b; Knight et al. 2017), are added to an AA-PE-EB lure, these multicomponent lures (AA-PE-EB-PAN-PET) remain highly attractive to codling moth and S. myopaeformis. This five component lure catches a suite of leafrollers (Lepidoptera: Tortricidae) including, Choristoneura rosaceana Harris, Pandemis limitata (Robinson), and Spilonota ocellana (Denis and Schiffermüller) and it remains as attractive to these leafrollers as their individual binary kairomones, AA-PET or AA-PAN, respectively. In this areawide SIT system, there may be few advantages to adding any species specific sex pheromones to these kairomones because other than codling moth, catches of females from each species are reduced when their own sex pheromone is combined with these kairomones. These multicomponent kairomones may aid development of a multispecies areawide pest management program in British Columbia by making mass trapping far more economical because material and labour pest control costs are distributed across several species. It

PheroFip 2019 36

remains to be seen whether these multicomponent lures can be produced reliably and economically by commercial enterprises.

Key words: apple, acetic acid, pear ester, phenylacetonitrile, 2-phenylethanol, ethyl butyrate

Acknowledgements We would like to thank Mr. Mark Gardiner, Summerland Research and Development Centre, and Mr. Bill Lingren, Trécé Inc., Adair, OK, USA, for making or developing the experimental lures used extensively in these trials.

References Judd, G. J. R., Knight, A. L. & El-Sayed, A. M. 2017a: Development of kairomone-based lures and traps targeting female Spilonota ocellana (Lepidoptera: Tortricidae) in apple orchards treated with sex pheromones. Canadian Entomol. 149: 662-676. Judd, G. J. R., Knight, A. L. & El-Sayed, A. M. 2017b: Trapping Pandemis limitata (Lepidoptera: Tortricidae) moths with mixtures of acetic acid, caterpillar-induced apple- leaf volatiles, and sex pheromone. Canadian Entomol. 149: 813-822. Knight, A. L., El-Sayed, A. M., Judd, G. J. R. & Basoalto, E. 2017: Development of 2- phenylethanol plus acetic acid lures to monitor obliquebanded leafroller (Lepidoptera: Tortricidae) under mating disruption. J. Appl. Entomol. 141: 729-739.

PheroFip 2019 37

Evaluating the trap efficiency of two pheromone compounds of male bean flower thrips in the field in Kenya

Falko P. Drijfhout2, David K. Mfuti1, Adeyemi O. Akinyemi1, Amanuel Tamiru1, Sevgan Subramanian1, Tom W. Pope3, Heather Campbell3, William D. J. Kirk2

1International Centre of Insect Physiology and Ecology, Nairobi, Kenya; 2Keele University, Staffordshire, UK 3Harper Adams University, Shropshire, UK

Abstract The bean flower thrips (BFT), Megalurothrips sjostedti (Thysanoptera: Thripidae), is a key pest of legumes in sub-Saharan Africa, especially cowpea, Vigna unguiculata (L.) Walp. and French bean, Phaseolus vulgaris L. (Fabaceae). Cowpea is important in subsistence farming and it is consumed daily by about 200 million people worldwide. Thrips are difficult to control because they are very small and retreat into small inaccessible spaces on plants. It causes abscission of cowpea flowers resulting in significant yield losses between 20 and 100%. A heavy infestation can cause total crop loss. To control the damage by BFT, farmers often rely on the use of pesticides. Overuse of these chemical products leads to detrimental consequences to the environment, e.g. leading to very high levels of insecticide residues on produce. Recently, two distinct pheromone compounds have been identified in the headspace of male BFT. Subsequent behavioural assays in the laboratory indicated enhanced attraction of both BFT males and females to these identified compounds. The exact role of the two compounds thus far remains unclear. Experiments using blue traps carried out in Kenya indicated that these compounds attract both males and females. There were significant differences in the attraction of female BFT between the different concentrations for both of the synthetic BFT pheromone compounds. In the presentation the differences in trap catches from the two compounds and their synergy will be presented and discussed. In addition to discussing the usefulness of these compounds in integrated pest management, the role of non- volatile hydrocarbons in mate and species recognition will be touched upon.

PheroFip 2019 38

Improving Grapholita molesta monitoring in peach and nectarine orchards under Mating Disruption by using bisexual lures

Michele Preti1*, Alan L. Knight2, Sergio Angeli1

1Faculty of Science and Technology, Free University of Bozen-Bolzano, Bolzano, Italy 2USDA, Agricultural Research Service, Wapato, Washington *[email protected]

Abstract Italy is together with Spain the leader producer of peaches and nectarines in Europe. Emilia- Romagna is the second most important Italian region, after Campania, hosting about 12% and 36% of the total peaches and nectarines surface, respectively. Such stone fruits are still key crops for the farms of this region, although in the last years a lower economic profit has been registered, with a consequent surface reduction. Peach and nectarine orchards in Emilia- Romagna are all managed with the Integrated Fruit Production (IFP), which includes the use of sex pheromones for Mating Disruption (MD) of oriental fruit moth, Grapholita molesta Busck (Lepidoptera: Tortricidae). In particular, G. molesta MD represents one of the best result achieved in insect pest control with pheromones. G. molesta MD is used by almost every farmer, combining it with few insecticidal interventions applied only in case of need and according to the G. molesta forecasting model of Emilia-Romagna Plant Protection Service. Unfortunately, during the last few years the occurrence of sporadic infestations of G. molesta has been registered also under MD control, alarming growers and local authorities. The occurrence of G. molesta damage in mating disrupted orchards, both on shoots and on fruits at harvest, have been recorded since 2016 only in some areas of Ravenna and Forlì- Cesena provinces. Even if these records are still limited to specific farms, this phenomenon is of great concern and several hypotheses have been considered. For example, today production area is halved compared to ten years ago and the average orchard size under MD may be too small; the timing of MD dispenser application may be too late compared to the phenology of the target pest; and perhaps growers are not supplementing MD with sufficient use of insecticides. Monitoring G. molesta flight under MD is not possible using the classical sex pheromone baited traps. Even if some farmers usually place few sex pheromone traps in the orchard borders to verify the effectiveness of the MD technique, the lack of catches does not always correspond to lack of presence. To overcome this issue, new kairomonal lures have been developed to be used under MD. Such tools, available since a few years for Cydia pomonella (L.), have been recently industrialized also for G. molesta. The aim of this work was to validate OFM Combo Dual® lure, a new commercial bait of Trécé (Adair, OK, USA), launched in the Italian market since 2018 by Certis Europe (Saronno, VA, Italy) to monitor G. molesta. OFM Combo Dual® is a similar lure of DA Combo®, a bisexual lure developed by Trécé to monitor C. pomonella using its sex pheromone boosted by the kairomone ‘Pear Ester’ or (E,Z)-2,4-ethyl decadienoate. OFM Combo Dual® exploits the attractiveness of different sexual and kairomonal volatile compound blends developed in the recent years. It includes a septa lure loaded with a blend of the three components of G. molesta sex pheromone and the major component of C. pomonella sex pheromone and a second membrane lure loaded with terpinyl acetate and acetic acid that allows traps to catch both moth sexes (Mujica et al. 2018).

PheroFip 2019 39

Trapping was conducted in Emilia-Romagna in 13 commercial orchards treated with several types of MD to compare moth catches in traps baited with either the sex pheromone Trécé OFM L2® lure versus the new kairomonal lure, Trécé OFM Combo Dual®. Five replicates of each lure type were randomized within each orchard using Trécé Delta VI traps® and white sticky liners. Each plot was minimum 1.5 hectares and the trap distance was minimum 25 m from the orchard border and between traps. Either white or orange delta traps were used in the different orchards and previous studies have found that they do not differ in the catch of G. molesta, but we wanted to evaluate any potential effect of trap color on the nontarget catch of pollinators. Each trial was performed for a period of 6-8 weeks during 2018. Our results showed that the new Trécé lure (OFM Combo Dual®) is significantly more effective than a standard sex pheromone lure to monitor G. molesta flight under MD, and is very effective in catching female moths. We also noted that the use of orange traps significantly reduced the catch of pollinators, i.e. honeybees and mason bees, which were never caught in orange traps. Finally, it was surprising to observe that male moths were caught also in the standard pheromone baited traps in some orchards despite the use of MD. The availability of new tools such as OFM Combo Dual® lure for both G. molesta males and females can improve monitoring of G. molesta under MD and help to bring this pest under better control.

Key words: Prunus persica, Lepidoptera, Tortricidae, delta trap, female moth, kairomones

References Mujica, V., Preti, M., Basoalto, E., Cichon, L., Fuentes-Contreras, E., Barros-Prada, W., Krawczyk, G., Nunes, M. Z., Walgenbach, J. F., Hansen, R. & Knight, A. L. 2018: Improved monitoring of oriental fruit moth (Lepidoptera: Tortricidae) with terpinyl acetate plus acetic acid membrane lures. Journal of Applied Entomology. 142: 731-744.

PheroFip 2019 40

Herbivory-induced plant volatiles from apple attract Archips xylosteana (Lepidoptera: Tortricidae)

Zaid Badra1, Marco Tasin2, Sergio Angeli1

1Faculty of Science and Technology, Free University of Bozen-Bolzano, Bolzano, Italy 2Department of Plant Protection Biology, Swedish University of Agricultural Sciences, Alnarp, Sweden

Abstract Insecticides are used by apple growers to control leafrollers. However, due to their negative impact on both human health and the ecosystem, new alternative tools are needed. In our previous studies, we have shown that specific bland of herbivore-induced plant volatiles (HIPVs) released by apple foliage are attractive to conspecific adult male and female of Pandemis heparana and Pandemis pyrusana (Lepidoptera: Tortricidae) when used together with acetic acid. In this research, we aimed to evaluate the effectiveness of several blends of herbivory-induced plant volatiles previously characterized in apple trees with and without acetic acid as a binary lure to monitor tortricid moths. In 2018, a field experiment was carried out in an organically managed apple orchard of South Tyrol (Italy) with the aim to assess several formulations of HIPVs as moth attractants. Ten blends of different HIPVs were loaded inside Eppendorf vials at different concentrations and placed in white delta traps. Eight replicates were randomized and spaced 6 m apart along tree rows. Traps were hung in the tree canopy at a 2 m height. Experiments started in late spring and lasted until August. Traps were checked once a week and lures were replaced every two weeks. Statistical analysis showed that all the treatments with acetic acid significantly caught more A. xylosteana than treatments without acetic acid. A. xylosteana is a polyphagous pest of fruit trees such as apple, pear, plum, and cherry. The larvae of A. xylosteana can cause significant damage by feeding on foliage and buds of apples trees and of other species. The majority of the caught individuals were males. The highest number of A. xylosteana moths was caught on June 28, with a mean of 22 moths per trap per week. To the best of our knowledge, the results of the present study provide the first effective lure for A. xylosteana. In agreement with our previous studies, we proved that specific lures of herbivory-induced plant volatiles from apple foliage can be used to monitor tortricid moths such as A. xylosteana when combined with acetic acid.

Key words: kairomone, monitoring, trapping, integrated pest management, Malus domestica

PheroFip 2019 41

SESSION 3

ALLELOCHEMICAL-BASED PEST

MANAGEMENT TACTICS

CHAIRPERSON: MICHELLE FOUNTAIN

PheroFip 2019 42

Allelochemicals, how useful are they in plant protection?

Olle Anderbrant1*

1Department of Biology, Lund University, Sölvegatan 37, SE-223 62 Lund, Sweden, *[email protected]

Abstract Long-distance sex or aggregation pheromones are common in many insect groups. They are often essential for mate finding and reproduction and therefore provide an obvious target for exploitation aiming for monitoring or controlling pest insects. However, there are also many species that rely on other cues to find mates and often these cues are weaker or less obvious to the researchers. The habitat or food sources may emit visual or olfactory signals guiding the insect to places they can mate and reproduce. Over the years we have investigated a number of systems where long-distance pheromones do not seem to be present, or where the use of non-pheromone lures would facilitate evaluation of pest control strategies. Here I give three examples where future research may lead to improved pest control using cues from hosts or food, including pests on stored food, vegetable crops and field crops. For a number of stored product pests useful pheromone-based monitoring or mating disruption (MD) strategies exist (Ryne et al. 2007). However, when evaluating MD effectiveness usually some non-pheromone based method should be used, and to measure product damage is often difficult in processing industries (Anderbrant et al., 2009). For Ephestia cautella water traps seem to give a reliable estimate of the relative populations (Ryne et al. 2006), whereas for Plodia interpunctella and potentially Ephestia kuehniella lures of volatile food compounds may provide a solution (Olsson et al. 2006). Carrots are grown in many parts of the world and are threatened by a number of herbivores. In parts of northern and central Europe, the carrot psyllid Trioza apicalis is the main pest and causes curling of leaves, deformation of the roots and also spread Liberibacter disease (Munyaneza et al. 2012). The psyllid antennae respond to compounds from the carrot as well as from conifers where they overwinter (Krisoffersen et al. 2008), but no long- distance odour-based behaviours have been observed. However, an earlier unidentified volatile carrot compound seems to influence the choice of host plant when psyllids can choose among a selection of carrot varieties containing different amounts of this compound. Could this be used in for instance push-pull strategies or to guide breeding programs? Finally, clover is an increasingly important crop, both for green manure and feed production, but clover seeds are often problematic to produce in large enough quantities due to an array of pests, especially the specialized clover seed weevils, Protapion (Apion) sp. (Lundin et al., 2017). Different weevil species are highly specialized and even if fields with different clover species are close to each other nearly 100% of the weevils in white clover are P. fulvipes and in red clover P. trifolii (Nyabuga et al. 2015). Clover odour profiles from different species overlap to a very large extent and different weevil species’ response profiles are almost identical (Andersson et al. 2012). How do the weevils manage and can we make use of their ability?

Key words: Semiochemicals, allomones, kairomones, food odour, Ephestia, Plodia, Trioza, Protapion

PheroFip 2019 43

Acknowledgements I would like to thank all co-authors and other collaborators that have contributed to the preliminary findings reported here, as well numerous funding agencies for their support.

References Anderbrant, O., Ryne, C., Sieminska, E., Svensson, G.P., Olsson, P.-O.C., Jirle, E. & Löfstedt, C. 2009: Odour signals for detection and control of indoor pyralid moths. IOBC wprs Bulletin 41: 69-74. Andersson, M. N., Larsson, M. C., Svensson, G. P., Birgersson, G., Rundlöf, M., Lundin, O., Lankinen, Å. & Anderbrant, O. 2012: Characterization of olfactory sensory neurons in the white clover seed weevil, Apion fulvipes (Coleoptera: Apionidae). Journal of Insect Physiology 58: 1325-1333. Kristoffersen, L., Larsson, M. C. & Anderbrant, O. 2008: Functional characteristics of a tiny but specialized olfactory system: olfactory receptor neurons of carrot psyllids (Homoptera: Triozidae). Chemical Senses 33: 759-769. Lundin, O., Svensson, G. P., Larsson, M. C., Birgersson, G., Hederström, V., Lankinen, Å., Anderbrant, O. & Rundlöf, M. 2017: The role of pollinators, pests and different yield components for organic and conventional white clover seed yields. Field Crops Research 210: 1-8. Munyaneza, J. E., Sengoda, V. G., Stegmark, R., Arvidsson, A. K., Anderbrant, O., Yuvaraj, J-K., Rämert, B. & Nissinen, A. 2012: First report of “Candidatus Liberibacter solanacearum” associated with psyllid-affected carrots in Sweden. Plant Disease 96: 453. Nyabuga, F. N., Carrasco, D., Ranåker, L., Andersson, M. N., Birgersson, B., Larsson, M. C., Lundin, O., Rundlöf, M., Svensson, G. P., Anderbrant, O. & Lankinen, Å. 2015: Field abundance patterns and odor-mediated host choice by clover seed weevils, Apion fulvipes and A. trifolii (Coleoptera: Apionidae). Journal of Economic Entomology 108: 492-503. Olsson. P.-O. C., Anderbrant, O. & Löfstedt, C. 2006: Attraction and oviposition of Ephestia kuehniella induced by volatiles identified from chocolate products. Entomologia Experimentalis et Applicata 119: 137-144. Ryne, C., Ekeberg, M., Jonzén, N., Oehlschlager, C., Löfstedt, C. & Anderbrant, O. 2006: Reduction in an almond moth Ephestia cautella (Lepidoptera: Pyralidae) population by means of mating disruption. Pest Management Science 62: 912-918. Ryne, C., Svensson, G. P., Anderbrant, O. & Löfstedt, C. 2007: Evaluation of long-term mating disruption of Ephestia kuehniella and Plodia interpunctella (Lepidoptera: Pyralidae) in indoor storage facilities by pheromone traps and monitoring of relative aerial concentrations of pheromone. Journal of Economic Entomology 100: 1017-1025.

PheroFip 2019 44

Interfering host location of Cacopsylla pruni with repellent plant volatiles

Jannicke Gallinger1,2, Cornelia Dippel3, Jürgen Gross1,2

1Julius Kühn-Institut, Federal Research Centre for Cultivated Plants, Institute for Plant Protection in Fruit Crops and Viticulture, Laboratory of Applied Chemical Ecology, Schwabenheimer Str. 101, Dossenheim, Germany; 2Technical University of Darmstadt, Chemical Plant Ecology, Schnittspahnstr. 10, Darmstadt, Germany; 3IS Insect Services GmbH, Berlin, Germany

Abstract The phloem-feeding plum psyllid Cacopsylla pruni is the only known vector of the phytoplasma ‘Ca. Phytoplasma prunorum’. This specialized cell wall-less bacterium is restricted to sieve elements in the phloem tissue of Prunus ssp.. Ca. P. prunorum causes European Stone Fruit Yellows (ESFY), one of the most severe diseases in stone fruits. Because infected Prunus cultivars yield poorly and die within a few years, ESFY is of high economic interest. C. pruni acquires the bacteria during feeding on infected plants. By transmission feeding the pathogens are spread to healthy Prunus trees. To date no effective control strategies or cures for phytoplasma diseases are available. To reduce the number of new infections and stem the spread of ESFY innovative control strategies against the vector are required. C. pruni is alternating its host plant two times within one generation (Ossiannilsson, 1992). C. pruni needs to reproduce on Prunus because nymphs are not able to develop on their second host coniferous trees (Gallinger & Gross, 2018). Therefore, in early spring adults arrive in stone fruit orchards for mating and ovipositing on Prunus spp.. After passing five nymphal stages the young adults (emigrants) migrate during summer to firs and other conifers to ensure adequate nutrition for the rest of the year until they return (remigrants) in early spring for reproduction to Prunus (Gallinger & Gross, 2018). The long distance alternation between these diverging plant species makes reliable host detection essential. Insects use secondary plant metabolites such as volatile organic compounds for localization of feeding and oviposition hosts. The use of olfactory cues for host location is shown for several psyllid species (Alquézar et al. 2017; Martini et al. 2014; Mas et al. 2014; Mayer et al. 2008a, 2008b, 2011; Mayer & Gross, 2007; Soroker et al. 2004). Interestingly a change in preference for volatiles from overwintering and reproduction hosts was found in the migrating psyllid Cacopsylla melanoneura (Mayer & Gross, 2007). Additionally the infestation of carrot plants with Trioza apicalis can be reduced by the application of sawdust from evergreen overwintering hosts (Nehlin et al. 1994). These findings underlie our hypothesis that C. pruni is attracted or repelled by specific volatiles from Prunus trees and conifers. We analyzed and compared the volatile profiles of Prunus trees and silver firs to identify species-specific components. In field surveys different infestation levels with C. pruni in Prunus varieties were found (Carraro et al. 2002; Mergenthaler et al. 2017). Therefore, additionally components that lead to the differentiation of volatile profiles from a high attractive Prunus rootstock and a less attractive cultivar were identified.

PheroFip 2019 45

Figure 1: Number of C. pruni remigrants which choose the pure odor of a Prunus tree (left) over the odor of a Prunus tree equipped with different repellent mixtures (green = Mix A –5%, red = Mix B – 5%, pale blue = Mix C–5%, mid-blue = Mix C–20%) incorporated in polypropylene plastic card dispensers (right). Significant differences are marked with asterisk (sign test, *P<.05, **P<.01, **P<.001).

Typical volatiles of the two Prunus varieties and silver firs were tested for antennal detection of C. pruni females and detectable volatiles were investigated for influence on psyllid behavior in Y-shaped olfactometer trails. Most compounds had a repellent effect on C. pruni adults. Mixtures of these repellent plant volatiles were incorporated in polypropylene plastic cards as dispensers. Their ability to mask the odor of an attractive host plant was verified in olfactometer trails (Fig. 1). Additionally these mixtures of potential repellents against C. pruni were tested in a first field study. An artificial application of those mixtures could be used for the development of an environmentally safe control strategy.

References Alquézar, B., Volpe, H. X. L., Magnani, R. F., Miranda, M. P. de, Santos, M. A., Wulff, N. A; Bento, J. M. S; Parra, J. R. P, Bouwmeester, H & Peña, L. 2017: β-caryophyllene emitted from a transgenic Arabidopsis or chemical dispenser repels Diaphorina citri, vector of Candidatus Liberibacters. Scientific reports 7: 5639. doi:10.1038/s41598-017-06119-w. Carraro, L., Ferrini, F., Ermacora, P. & Loi, N. 2002: Role of wild Prunus species in the epidemiology of European stone fruit yellows. Plant Pathology 51: 513–517. doi:10.1046/j.1365-3059.2002.00732.x.

PheroFip 2019 46

Gallinger, J. & Gross, J. 2018: Unraveling the Host Plant Alternation of Cacopsylla pruni - Adults but Not Nymphs Can Survive on Conifers Due to Phloem/Xylem Composition. Frontiers inPplant Science 9: 484. doi:10.3389/fpls.2018.00484. Martini, X., Kuhns, E. H., Hoyte, A. & Stelinski, L. L. 2014: Plant volatiles and density- dependent conspecific female odors are used by Asian citrus psyllid to evaluate host suitability on a spatial scale. Arthropod-Plant Interactions 8: 453–460. doi:10.1007/s11829-014-9326-z. Mas, F., Vereijssen, J. & Suckling, D. M. 2014: Influence of the pathogen Candidatus Liberibacter solanacearum on tomato host plant volatiles and psyllid vector settlement. Journal of chemical ecology 40: 1197–1202. doi:10.1007/s10886-014-0518-x. Mayer, C. J. & Gross, J. 2007: Different host plant odours influence migration behaviour of Cacopsylla melanoneura (Förster), an insect vector of the apple proliferation phytoplasma. IOBC/WPRS Bull. 30: 177–184. Mayer, C. J., Vilcinskas, A. & Gross, J. 2008a: Pathogen-induced release of plant allomone manipulates vector insect behavior. J. Chem. Ecol. 34: 1518–1522. doi:10.1007/s10886- 008-9564-6. Mayer, C. J., Vilcinskas, A. & Gross, J. 2008b: Phytopathogen lures its insect vector by altering host plant odor. J. Chem. Ecol. 34: 1045–1049. doi:10.1007/s10886-008-9516-1. Mayer, C. J., Vilcinskas, A. & Gross, J. 2011: Chemically mediated multitrophic interactions in a plant-insect vector-phytoplasma system compared with a partially nonvector species. Agric. For. Entomol. 13: 25–35. doi:10.1111/j.1461-9563.2010.00495.x. Mergenthaler, E., Kiss, B., Kiss, E. & Viczián, O. 2017: Survey on the occurrence and infection status of Cacopsylla pruni, vector of European stone fruit yellows in Hungary. Bull. Insectology 70: 171–176. Nehlin, G., Valterová, I. & Borg-Karlson, A. K. 1994: Use of conifer volatiles to reduce injury caused by carrot psyllid,Trioza apicalis, Förster (Homoptera, Psylloidea). Journal of chemical ecology 20: 771–783. doi:10.1007/BF02059612. Ossiannilsson, Frej (Hg.) 1992: The Psylloidea (Homoptera) of Fennoscandia and Denmark. Fauna entomologica Scandinavica 26. ed. Ossiannilsson, F. (Leiden, New York, Köln: Brill). Soroker, V., Talebaev, S., Harari, A.& Wesley, S. D. 2004: The role of chemical cues in host and mate location in the pear psylla Cacopsylla bidens (Homoptera: Psyllidae). Journal of Insect Behavior 17: 613-626. doi:10.1023/B:JOIR.0000042544.35561.1c

PheroFip 2019 47

Host defense chemicals as a repellent to Xylosandrus germanus ambrosia beetle infestations in apple trees

Arthur Agnello1, Kenneth Lamm2

1Cornell University, Department of Entomology, New York State Agricultural Experiment Station, Geneva, New York USA; 2Bowdoin College, Department of Biology, Brunswick, Maine USA

Abstract The black stem borer, (BSB, Xylosandrus germanus), is an ambrosia beetle that is a serious pest in ornamental tree nurseries and forests in the US. It is native to Asia and was first documented in NY in 1932. Current studies indicate that it invades orchards from nearby wooded areas. The insects attack stressed (including some ‘apparently healthy’) trees, boring into the trunk or limbs to create galleries where young develop. The trees wall off their vascular system in response to the attack, causing tree decline and death. Infestations in commercial apple orchards were seen for the first time in 2013, in multiple western NY sites, causing up to 30% tree loss in some sites. In our initial trials, we used traps baited with ethanol (an attractant for these beetles) to document the extent and timing of their occurrence in over 50 NY orchards. We have also tested various insecticide sprays for their control and investigated the use of a repellent, verbenone, applied as either a volatile or topically applied formulations to deter beetle attacks. While none of the currently available products provided acceptable levels of control, our trials in 2017 also included an experimental formulation of verbenone combined with methyl salicylate, another known plant defense compound produced by plants under stress; this combination resulted in zero BSB attacks or infestations in our trials, and therefore represented a possibility for further testing and evaluation in the field. Material and Methods In 2018, we tested trunk applications of different repellents for X. germanus control in potted apple trees, which were waterlogged to stress them to produce ethanol, and placed inside wooded areas directly adjacent to orchard sites with documented X. germanus infestations. Additionally, individual ethanol lures were attached to each tree to increase their attractiveness to the beetles. The potted apple trees placed were deployed in three different apple orchards in Wayne County, NY, USA. SPLAT flowable wax matrix (ISCA Technologies [Riverside, CA]) containing different combinations of verbenone and methyl salicylate repellents, was applied at different rates on the apple tree trunks. Other treatments tested were chlorpyrifos (Lorsban, Dow AgroSciences [Indianapolis, IN]), and acibenzolar-S- methyl (Actigard, Syngenta [Greensboro, NC]), a chemical used to promote of the tree's defense mechanisms. The untreated trees were either flood-stressed or drought-stressed. Trees were removed on 10 July, after the end of the beetles' first flight, and destructively sampled for BSB infestations, to determine what effect these repellent treatments had in preventing attacks by this ambrosia beetle. Results Trees treated with the highest rate (35g per tree) of the verbenone+methyl salicylate formulation had no infestation holes across all sites. Also, there was no damage in the trees treated with acibenzolar-S-methyl. Chlorpyrifos-treated trees had the highest number of holes/trunk. Trees treated with the mixtures of verbenone and methyl salicylate had fewer holes/trunk than did trees treated with verbenone or methyl salicylate alone. The results indicate that a high rate of the SPLAT verbenone+methyl salicylate formulation would be

PheroFip 2019 48

most effective in preventing infestations, and that the application would need to be made before the first flight, sometime in late April or early May, before any BSB are actively flying. Further research of the optimal application periods would allow a better understanding of when and how repellents should be used for maximum efficacy against BSB attacks. Use of verbenone+methyl salicylate could minimize the need for broad-spectrum insecticides such as chlorpyrifos, which would promote better integrated pest management.

Key words: black stem borer, Xylosandrus germanus, apples, trunk sprays, verbenone, methyl salicylate

PheroFip 2019 49

Attractiveness of different lure and infect devices for thrips control

Robert W.H.M. van Tol1, Melanie M. Davidson2, Ruth C. Butler2, David A.J. Teulon2, Willem Jan de Kogel1

1Wageningen University and Research, P.O. Box 69, 6700 AB Wageningen, the Netherland 2Institute for Plant and Food Research Limited, Private Bag 4704, Christchurch, New Zealand

Abstract Lure and infect strategy for control of pest insects is considered as a pest management strategy with high potential, but so far only few have proven to be effective for application in agriculture. While more attractive semiochemicals and effective entomopathogenic fungi become available for pest control, visually attractive devices where the insects have to pick up the fungal spores are still poorly developed. We tested the efficacy in terms of number of thrips landing on differently coloured and structured trap types in order to find efficient traps that could be further developed into effective auto-dissemination devices. We found a high visual response to a sticky white ruffle trap (flexible 30cm long cylindrical folded fabric) compared to a cylindrical folded commonly used blue sticky plate product (Bug-scan®) for three thrips species, the western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), the onion thrips, Thrips tabaci (Lindeman) (Thysanoptera: Thripidae) and the New Zealand flower thrips, Thrips obscuratus (Crawford) (Thysanoptera: Thripidae) in the greenhouse (roses) and field (pasture). A similar blue coloured ruffle and the adapted Lynfield trap performed poorly compared to both white ruffle and the Bug-scan® plate. Addition of the thrips attractant methyl isonicotinate (MI) increased attraction of T. tabaci to the different trap types in the field. The white ruffle without MI was however always more attractive than the Bug-scan® (~3x more) or any of the other trap types tested without MI (11 to 22x more). A strong interaction between the white ruffle and MI was found for T. obscuratus in the field. All trap types tested with and without MI caught none or very few T. obscuratus (0 to 1.8 per trap) whereas the white ruffle with MI caught 15.4 thrips/trap (white ruffle without MI 0.9 trips/trap) suggesting a strong odour induced visual response. In the greenhouse only weak attractive effect of MI to the tested trap types was found for F. occidentalis. The white ruffle trap by itself was on average 3.5 (Bug-scan), 7 (blue ruffle) and 14x (Lynfield) more attractive with little or no effect of MI. We discuss the role of different visual aspects of the ruffle trap in the decision making of thrips to land on objects as well as the importance of visual factors for future design of more effective auto-dissemination traps for the control of thrips.

Key words: Frankliniella occidentalis, Thrips tabaci, Thrips obscuratus, Anaphothrips obscurus, Tenothrips frici, colour, methyl isonicotinate, semiochemical, Thysanoptera, Thripidae, monitoring, lure and infect

PheroFip 2019 50

Push-Pull with Synthetic Semiochemicals for Control of Fruit Pests

Michelle T Fountain 1, Chantelle Jay1, Adam Walker1, Francesco Rogai1, David Hall2, Dudley Farman2

1NIAB EMR, New Road, East Malling Kent ME19 6BJ 2Natural Resources Institute, University of Greenwich, Chatham Maritime, Kent ME4 4TB, UK

Abstract Insect pest species in fruit crops are controlled by a range of management strategies including cultural, biological and synthetically produced pesticides. Changes in insecticide approvals and climate change coupled with non-native species introductions are jeopardising current control practices and risk disrupting Integrated Pest Management. Push-pull strategies have an element which repels pests (the push), and an attractant source to draw pests away from the crop (the pull). The pull can be combined with a killing agent to prevent the pest re-entering the crop and to reduce population growth and, hence, crop damage. Using synthetic semio-chemicals, two push-pull systems were tested for control of Lygus rugulipennis and Drosophila suzukii. We investigated whether fruit damage could be reduced using synthetically produced repellents, attractants or a combination of the two (push-pull). In field trials with replicated plots (25 x 25 m) significantly fewer adult and nymph L. rugulipennis were present where the push or push-pull combined was applied. There was no significant effect of the pull treatment when used alone. There were also fewer damaged fruits where a ‘push’ treatment and a ‘pull’ treatment were combined compared to no treatment. Preliminary field trials on D. suzukii in cherry orchards indicated that some coded synthetic volatile substances reduced the numbers of eggs laid in fruits. Push–pull field trials are currently underway in strawberry crops and results will be presented.

Key words: attractants, capsid, mirid, push-pull, repellent, SWD, synthetic

PheroFip 2019 51

Sucked in! Crop borders to reduce aphid-transmitted viruses

Kerstin Krüger1*, Michelle L. Schröder1, Robert Glinwood, Rickard Ignell

1Department of Zoology and Entomology, University of Pretoria, Private Bag X20, Pretoria, 0028, South Africa; 2Department of Crop Protection Ecology, Swedish University of Agricultural Sciences, Uppsala, Sweden; 3Unit of Chemical Ecology, Department of Plant Protection Biology, Swedish University of Agricultural Sciences, Alnarp *[email protected]

Abstract Many studies have shown that plant volatile cues are important for aphids to locate their host plants (Bruce & Pickett, 2011). Yet, exploiting volatiles for the management of plant viruses through manipulating the behaviour of insect vectors is lagging behind. Plant borders consisting of a non-virus host plant around the edges of fields can reduce the incidence of non-persistently aphid-transmitted viruses (DiFonzo et al. 1996). Alatae (winged aphids) landing in plant borders purge their mouthparts of the virus when probing a virus non-host plant before moving into the field proper. The efficacy of crop borders in decreasing virus transmission is variable (Hooks & Fereres, 2006). For crop borders to be effective aphid landing rates need to be higher at the edges compared to the interior of a field (edge effect). Although edge effects were apparent in trials with 20 to 40 ha seed potato fields in South Africa, patterns within and among sites were not always consistent. In order to strengthen the edge effect, crops border plants should be more attractive than the main crop to aphid vectors. Identification of behaviourally active plant volatiles may aid in optimizing the selection of attractive crop border plants. Rhopalosiphum padi (bird-cherry oat aphid) and Macrosiphum euphorbiae (potato aphid) are two important vectors of the non-persistently transmitted Potato virus Y (PVY). The virus leads to economic losses in seed potato-growing regions in many parts of the world due to downgrading of seed potatoes. The responses of alate adult aphids to plant volatiles of three maize, wheat and potato cultivars each, were determined in no-choice bioassays using a four- arm olfactometer. Different cultivars of the same plant species were tested since they may differ in their attractiveness to aphids. The oligophagous R. padi, which colonizes Poaceae, was attracted to odours of one maize cultivar (‘6Q-121’). The polyphagous M. euphorbiae was neither attracted nor repelled by any of the cultivars tested. Macrosiphum euphorbiae colonizes plant species within different families and may therefore be less reliant on olfactory cues than R. padi. An analysis of headspace volatiles from each cultivar with coupled gas chromatography/mass spectrometry (GC-MS) revealed quantitative and qualitative differences. When determining the response of R. padi to individual compounds in olfactometer trials, (Z)-3-hexenyl acetate, which occurred in the highest concentration in maize cultivar ‘6Q-121’, attracted R. padi. Four compounds, α-farnasene, (E)-2-hexenal, indole and TMTT, which were recorded in several of the plant cultivars, repelled R. padi. Thus, maize ‘6Q-121’ which attracted R. padi and did not emit compounds repellent to this species, may be a suitable crop border plant for seed potato fields where R. padi is an important vector of PVY (Schröder et al. 2015). The selection of effective crop border plants is crucial for the management of aphid- transmitted viruses based on insect behaviour. Identification of plant volatiles combined with bioassays to evaluate the olfactory response in the laboratory can facilitate the pre-screening of potential crop border plants before proceeding with field trials. Crop border plants should

PheroFip 2019 52

not emit volatile compounds that repel aphids since they may reduce landing and subsequent probing (Schröder et al. 2017). Understanding the relationship between plant volatiles and aphid host plant selection behaviour is an important step in the successful development of crop borders to control the spread of non-persistent viruses such as PVY.

Key words: Aphididae, Hemiptera, host plant, olfaction, plant virus, plant volatiles, attractive, repellent

References Bruce, T. J. A. & Pickett, J. A. 2011: Perception of plant volatile blends by herbivorous insects – Finding the right mix. Phytochemistry 72: 1605-1611. DiFonzo, C. D., Ragsdale, D. W., Radcliffe, E. B., Gudmestad, N. C. & Secor, G. A. 1996: Crop borders reduce potato virus Y incidence in seed potato. Ann. Appl. Biol. 129: 289- 302. Hooks, C. R. R. & Fereres, A. 2006: Protecting crops from non-persistently aphid-transmitted viruses: A review on the use of barrier plants as a managed tool. Virus Res. 120: 1-16. Schröder, M. L., Glinwood, R., Ignell, R. & Krüger, K. 2017: The role of visual and olfactory plant cues in aphid behaviour and the development of non-persistent virus management strategies. Arthropod-Plant Interact. 11: 1-13. Schröder, M. L., Glinwood, R., Webster, B., Ignell, R. & Krüger, K. 2015: Olfactory responses of Rhopalosiphum padi to three maize, potato, and wheat cultivars and the selection of prospective crop border plants. Entomol. Exp. Appl. 157: 241-253.

PheroFip 2019 53

A new bisexual kairomone lure for codling moth

Alan L. Knight1, Valentina Mujica2, Marco Tasin3

1Yakima Agricultural Research Laboratory, USDA-ARS, 5230 Konnowac Pass Rd, Wapato, Washington 98951, USA. [email protected]; 2INIA, Instituto Nacional de Investigación Agropecuaria, Estación experimental Las Brujas, Ruta 48 km 10 Canelones 90200, Uruguay. [email protected] 2Oregon State University, 530 Hanley Rd., Central Point, Oregon 97502, USA. [email protected]; 3Integrated Plant Protection Unit, Department of Plant Protection Biology, Swedish University of Agricultural Science, 23053 Alnarp, Sweden. [email protected]

Abstract Finally, the promise of pear ester (E, Z)-2,4-decadienoate as a potent bisexual attractant for codling moth Cydia pomonella (L.) has been realized through the creation of powerful new multi-component kairomone blends. This discovery comes nearly 20 years after Dr. Douglas Light revealed the attractiveness of pear ester as the first identified individual kairomone that was effective at rates similar to a sex pheromone and attractive to both sexes of moths (Light et al. 2001). Pear ester used within walnut groves is a powerful attractant, but in pome fruits many factors impacted its effectiveness, such as crop, cultivar, seasonality, crop load, and the presence of fermenting fruit on the orchard floor. Pear ester also performed well when used in combination with the sex pheromone and a ‘Combo’ lure has been adopted throughout the world to monitor C. pomonella in orchards treated with sex pheromones for mating disruption (Knight et al. 2005). A number of studies have developed lure blends including pear ester that have improved its performance (Landolt et al. 2007; Landolt et al. 2014; Jaffe & Landolt, 2018). The use of n-butyl sulfide in combination with acetic acid and pear ester (BSAAPE) has been purported to be a more effective lure than pear ester alone and has recently been tested effectively to mass trap C. pomonella in commercial orchards (Jaffe et al. 2018). In a continuing effort to develop new bisexual lures for C. pomonella a series of field trials were conducted with delta traps with removable liners and with bucket traps using propylene glycol during 2018. During a serial process of testing new attractant blends a potent multi- component blend was identified. This blend consistently outperformed the use of BSAAPE, by 3-fold. The proportion of female moths caught with the new blend ranged from 60 to 80% during the summer. The new blend was equally effective in apple blocks with low fruit loads and in an orchard with a heavy fruit load with low (early season), moderate (mid-season) and high levels (late season) of fruit injury and even later in the season when the orchard floor was littered with fermenting fruits. The new blend outperformed several sex pheromone lures in blocks both untreated and treated with sex pheromone dispensers. The attractiveness of this blend was also tested in combination with sex pheromone lures and male catch was significantly increased and female catch declined slightly when the sex pheromone was added. Subsequent tests have expanded the list of plant volatiles that can be effective when substituted within this blend. Trials continue to develop these lures in the southern hemisphere during 2018-19 and updated information will be presented at the meeting.

Key words: apple, kairomones, n-butyl sulfide, acetic acid, pear ester

Acknowledgements We would all like to thank Bill Lingren, Trécé Inc., Adair, OK, for developing experimental lures used extensively in these trials

PheroFip 2019 54

References

Jaffe, B. D. & Landolt, P. J. 2018: Field experiment of a three-chemical controlled-release dispensers to attract codling moth (Cydia pomonella) (Lepidoptera: Tortricidae). J. Econ. Entomol. 111: 1268-1274. Jaffe, B. D., Guedot, C. & Landolt, P. J. 2018: Mass-trapping codling moth, Cydia pomonella (Lepidoptera: Tortricidae), using kairomone lure reduce fruit damage in commercial apple orchards. J. Econ. Entomol. 111: 1983-1986. Knight, A. L., Hilton, R. & Light, D. M. 2005: Monitoring codling moth (Lepidoptera: Tortricidae) in apple with blends of ethyl (E,Z)-2,4-Decadienoate and codlemone. Environ. Entomol. 34: 598-603. Landolt, P. J., Suckling, D. M. & Judd, G. 2007: Positive interaction of a feeding attractant and host kairomone for trapping the codling moth (Lepidoptera: Tortricidae). J. Chem. Ecol. 33: 2236-2244. Landolt, P. J., Ohler, B., Lo, P., Cha, D., Davis, T. S. Suckling, D. M. & Brunner, J. 2014: N- butyl sulfide as an attractant and coattractant for male and female codling moth (Lepidoptera: Tortricidae). Environ. Entomol. 43: 291-297. Light, D. M., Knight, A. L. Henrick, C. A., Rajapaska, D., Lingren, B., Dickens, J. C., Reynolds, K. M., Buttery, R. G., Merrill, G., Roitman, J. & Campbell, B. C. 2001: A pear derived kairomone with pheromonal potency that attracts male and female codling moth, Cydia pomonella (L.). Naturwissenschaften 88: 333-338.

PheroFip 2019 55

SESSION 4

PHEROMONE-BASED PEST

MANAGEMENT TACTICS

CHAIRPERSON: ALLY HARARI

PheroFip 2019 56

History of a Mating Disruption Mega-dispenser: MSTRS Aerosols and Baggies

T. C. Baker1

1Penn State University, University Park, PA, U.S.A.

Abstract Dispensers for emitting sex pheromone for pheromone mating disruption have increasingly been engineered to be deployed as high-release-rate, widely spaced dispensers. Such mega- dispensers have been effective at producing satisfactory population control and reducing damage to agricultural crops. Whereas attention with regard to mating disruption success has traditionally been paid to achieving emission rates in terms of amounts emitted per acre or hectare per day, our laboratory has focused on the amounts emitted per dispenser per minute. The key reason for this focus is because we and others learned through long series of behavioral experiments in the 1980s and 1990s that upwind flight responses of moths to pheromone occurs via the moths’ upwind surges in response to individual pheromone strands in pheromone plumes. Because upwind flight is due to the intensity of molecular flux of individual strands, we focused on producing dispensers that would create the strongest possible strands downwind. In 1993 we first started researching the efficacy of mating disruption mega-dispensers that were comprised of pressurized canisters producing metered aerosols of pheromone emitted onto cloth pads on a timed-release basis during certain hours of the day or night. We soon replaced the pressurized canister type of Metered Semiochemical Timed Release System (“MSTRS”) with a non-pressurized pump-spray bottle type that emitted the same amounts of aerosols as those emitted by the pressurized canisters. From these two aerosol systems we gained critical knowledge about the pheromone emission rates emanating from the pads that were needed in order to achieve successful mating disruption in the field. By carefully measuring the emission rates from the pads we also realized that the pads receiving the timed aerosol sprays quickly reached a saturation point, after which the sprays did not further increase the pads’ emission rates. In other words, within several days, the pads became totally passive dispensers that were very effective at achieving mating disruption. Following these discoveries, working with Cam Oehlschlager of ChemTica Internacional we came up with a new type of MSTRS dispenser that was a totally passive- release system: polyethylene “baggies” whose emission rates mimicked those of the pads. The MSTRS baggies have been shown to be effective mating disruption dispensers against a variety of moth pest species.

PheroFip 2019 57

Insect pheromones in Portugal for crop protection: history and present situation

Carlos Frescata1

1BIOSANI, Quinta de S. Brás, Serra do Louro, 2950-354 Palmela, Portugal [email protected]

Abstract During the 1970s started in Portugal the use of insect sex pheromones for crop protection. The crop protection authorities (former CNPPA), namely through the forecast and warning services, started to use it for monitoring the main lepidopteran pest species. Regarding their use as a control measure during the 1980s happen the first trials of mating disruption for Cydia pomonella based in RAK. The 1990s was the decade of the expansion of pheromone lures for monitoring and the introduction of mating disruption in practice. This last one based in Shin-Etsu dispensers: Isomate C for C. pomonella and Isonet L for Lobesia botrana (Frescata, 1999ab, 2000ab). Madeira-Med Programme regarding Ceratitis capitata control (Pereira et al 2000) and the recently created IPM associations namely of pome fruit and grape growers were the main reasons for the impressive growth in the use of monitoring pheromone and parapheromone lures during this decade. The first decade of this century was clearly marked by the expansion of mating disruption in a wider use (Frescata, 2003, 2004). Douro wine region, through the farmers association ADVID, had the ambition to generalize it (Carlos et al. 2005). The use of mass trapping for C. capitata, through parapheromenes and food attractants, had a significant expansion. Two main innovative research projects appeared: Isonet CFM – creation based in Portuguese research of a new mating disruption dispenser for Prays citri control (Silva et al. 2006); mass trapping of Monochamus galloprovincialis, vector of the pine wood nematode with pine allelochemicals (Bonifácio et al. 2012). Present decade of 2010 it was marked by an effort trying to start the creation of new biotechnological tools based in differentiating scientific knowledge and resources available in the country. Using nanotechnological components started the conception of a sprayable formulation with Planococcus ficus sex pheromone to kairomonally attract in the vineyard its natural enemy Anagyrus spec. nov. near pseudococci and simultaneously having mating disruption effect (Mourato et al. 2014). This new control measure research was based in an earlier chemical ecology research in Portugal of Planococcus citri and P. ficus also including the mentioned parasitoid (Franco et al 2008). A new pheromone monitoring lure with nanotechnological components was created and patented, aiming to offer a more constant pheromone release (Silva & Frescata, 2014; Petersen-Silva et al. 2014). Nowadays, close to 2020 decade, the use of mating disruption covers an area of around 4000 ha, namely for L. botrana and C. pomonella control, and are already registered in Portugal nearly all the mating disruption products available in Europe.

Key words: mating disruption, monitoring, Cydia pomonella, Lobesia botrana, Prays citri, Planoccocus ficus, Monochamus galloprovincialis, nanotechnology

PheroFip 2019 58

References

Bonifácio, L., Praias. F. & Sousa, E. 2012: Trapping Monochamus galloprovincialis (Coleoptera: Cerambycidae), vector of the pine wood nematode, with pine allelochemicals, in Portugal. Silva Lusitana 20 (1/2): 39-53. Carlos, C., Costa, J., Gaspar, C., Domingos, J., Alves, F. & Torres, L. M. 2005: Mating disruption to control grapevine moth, Lobesia botrana (Den. & Schiff.), in Porto wine region: a three-year study. IOBC-WPRS Bull. 28 (7): 283-287. Franco, J. C., Silva, E. B., Cortegano, E., Campos, L., Branco, M. R., Zada, A. & Mendel, Z. 2008: Kairomonal response of the parasitoid Anagyrus spec. nov. near pseudococci to the sex pheromone of the vine mealybug. Entomologia Experimentalis et Applicata 126:122- 130. Frescata, C. 2000a: Mating disruption pheromones release in Portugal. XXI International Congress of Entomology, Foz do Iguaçu, Brasil, Ag. 2000, Vol. I: 169. Frescata, C. 2000b: Mating disruption situation in Portugal. 25th Anniversary Jubilee Meeting of the IOBC-WPRS Working Group “Use of Pheromones and Other Semiochemicals in Integrated Control”, Samos, Greece, September 25-29, 2000 (Abstracts): 25. Frescata C. 2003: Confusão sexual contra pragas em Portugal: superfícies, viabilidade e dificuldades. VI Encontro Nacional de Protecção Integrada, Castelo Branco, Maio 2003: 444-447. Frescata, C. 2004: Limitação de Pragas Sem Luta Química. Publicações Europa-América, Mem Martins, Portugal. Frescata, C., Gomes, C., Aires, A., Oliveira, M. & Assunção, A. 1999a: Confusão sexual para limitação de Lobesia botrana. V Encontro Nacional de Protecção Integrada, Bragança, Out. 1999: 73-79. Frescata, C., Gonçalves, M., Parente, P., Dias, L. & Bernardes, P. 1999b: Confusão sexual para limitação de Cydia pomonella em pomóideas. V Encontro Nacional de Protecção Integrada, Bragança, Out. 1999: 141-147. Mourato, C., Petersen-Silva, R., Neto, R., Franco, J. C. & Frescata, C. 2014: New perspectives on the biological control of Planococcus ficus. [WWW document] URL https://www.abim.ch/fileadmin/abim/documents/presentations2014/3_Frescata_Carlos_5 _ABIM2014.pdf. Cited 16 Nov. 2018. Pereira, R., Barbosa, A., Silva, N., Caldeira, J., Dantas, L. & Pacheco, J. 2000: Madeira-Med, a sterile insect technique programme for control of the Mediterranean fruit fly in Madeira, Portugal. In: Area-wide Control of Fruit Flies and Other Insect Pests, ed. Tan: 433-438. Petersen-Silva, R., Silva, J., Eira, R., Bonifácio, L., Vieira, F. & Frescata, C. 2014: New nano-developed pheromone lure for Cydia pomonella monitoring. [WWW document] URL:https://www.abim.ch/fileadmin/abim/documents/presentations2014/2_Frescata_Car los_6_ABIM2014.pdf . Cited 16 Nov. 2018. Silva, E. B., Gaspar, R., Dias, L., Antunes, R., Lourenço, I., Clemente, J. & Franco, J. C. 2006: Developing a mating disruption tactic for pest management of citrus flower moth. IOBC-WPRS Bull. 29(3): 127-137. Silva, J. & Frescata, C. 2014: Development of an innovative monitoring pheromone dispenser. IOBC-WPRS Bull. 99: 149-151.

PheroFip 2019 59

Studies on Lobesia botrana disruption mechanisms based on response after pheromone exposition

Aitor Gavara1, Sandra Vacas1, Jaime Primo1, Vicente Navarro-Llopis1

1CEQA-Instituto Agroforestal del Mediterráneo, Universitat Politècnica de València, Camino de Vera s/n, edificio 6C-5ª planta, 46022 Valencia (Valencia), Spain

Abstracts Mating disruption is a successful tool to control and maintain Lobesia botrana (Denis & Schiffermüller) (Lepidoptera: Tortricidae) populations under the economic threshold; nevertheless, the disruption mechanism acting in this case is not well known. Laboratory experiments were carried out in order to understand the mechanisms by which the disruption is triggered and to obtain the airborne pheromone concentration needed to provoke the desired effect. In these experiments, laboratory-reared L. botrana males were kept in glass chambers with a measured air stream and a known pheromone concentration. For this purpose, the pheromone emitted by a commercial mating disruption dispenser carried by the air stream was pushed through the glass chamber containing males to expose the individuals to pheromone concentration at different time intervals. A Porapak-Q cartridge was connected to the exit of the glass chamber to collect and quantify the pheromone emitted at known intervals. Different dosages were tested by exposing males to pheromone during 1, 3 and 24 hours. The effect of these dosages on male behavior was tested 24 hours after exposure. Electroantennography (EAG) tests were performed with exposed males to detect changes in pheromone detection. Likewise, wind tunnel tests were carried out to detect changes in male behavior. All these tests allowed us to elucidate the quantity of pheromone needed to produce disruption and to compare these values with existing field concentrations. Moreover, this information will help to understand which disruption mechanism is acting at each pheromone concentration.

Key words: European grapevine moth; Lepidoptera; mating disruption; sex pheromone

PheroFip 2019 60

Management of the honeydew moth by mating disruption in vineyard

Patricia Acín1

1Sociedad Española de Desarrollos Químicos S.L. (SEDQ), Marie Curie 33, 08210 Barberà del Vallès, Barcelona, Spain [email protected]

Abstract Cryptoblabes gnidiella is a Lepidopteran species, which has increased noticeably in the last few years in different crops in the South of Europe, especially in pomegranate, kaki and vineyards. The scarcity of products for its effective control along with the extended use of mating disruption for other coexisting species, such as Lobesia botrana in vineyards, makes crucial to develop an alternative based on the use of pheromones for the management of this species. Therefore, the efficacy of mating disruption with the dispenser CRYPTOTEC, at two different doses of dispensers per hectare, was tested in a plot of vineyards in the south of France. The results accomplished so far, show the effectiveness of the dispensers tested, and therefore the mating disruption as a possible strategy in the management of this species.

Introduction The honeydew moth, Cryptoblabes gnidiella Millière (Lepidoptera: Pyralidae) is a polyphagous species native to the Mediterranean region which has spread to different areas of Africa, Asia, Europe, New Zealand, Hawaii and South America. It has been mainly related to citrus, pomegranate and grapes, although it has been found in several other crops such as avocado, loquat and kaki. In Southern Europe it has been considered as a secondary pest but in the last years its population and correlated damage has increased considerably in certain crops such as kaki, pomegranate and vineyard. The efficacy of mating disruption in the control of C. gnidiella has been checked in the last 4 years in pomegranate and kaki (Acín, 2018). In vineyard, most of the studies have been focused mainly on the monitoring of this species (Anshelevich et al., 1993, Vidart et al., 2013, Demirel, 2016, Öztürk, 2018), but just a few experiencies in mating disruption has been performed, although not concluding enough. Therefore, the efficacy of mating disruption has been evaluated in a plot of vineyard with a high pest pressure, in order to assess the effect of the dispensers tested. This strategy entails a promising tool for the management of this species.

Material and methods A trial was performed in Baho, in the department of Pyrénées-Orientales in France. Two plots of 2.3 ha were treated with the mating disruption dispenser, CRYPTOTEC, at two different doses, 300 and 400 dispensers per hectare. A third plot of 2 ha approximately, without pheromone dispensers, was considered as control. The variety found in all plots was Grenache noir. These plots were chosen due to the high presence of G. gnidiella in the last years. Dispensers were placed before the first flight of adults following a homogeneous distribution. Each dispenser was comprising a blend of Z11-16:Ald and Z13-18:Ald in a ratio 1:1 and a maximum load of 330 mg of pheromone blend. Four delta traps with the corresponding monitoring dispenser were placed in each plot to monitor this species. These dispensers were replaced every 35-45 days.

PheroFip 2019 61

All 4 monitoring traps per plot were evaluated at least once a week considering the number of males caught in the devices. The efficacy of mating disruption was also considered by the evaluation of damages. For this purpose, an assessment of damage was accomplished just before harvest.

Results and discussion The number of captures found in the monitoring traps placed in the control was significantly higher than in both plots with mating disruption, with a total of 1713 captures compared to 1 and 3 in the plots with 400 dispensers/ha and 300 dispensers/ha. Furthermore, a patent difference was observed between the captures recorded in the two traps located close to the mating disruption plots and the two situated at a greater distance. An average of 623.5 captures were recorded in the two monitoring traps located further from the two plots in mating disruption and 233 in the traps situated closer. Regarding the efficacy in terms of damage, significant differences were found between treatments, being the control the plot with the highest percentage of damages with 17% of damaged bunches in comparison to 4% and 6.5% in mating disruption with 400 and 300 dispensers per hectare respectively. These results show a good efficacy of mating disruption in the control of C. gnidiella in vineyard at the two doses tested.

Key words: Honeydew moth, mating disruption, vineyard

References Acín, P. 2018: La confusión sexual como nueva herramienta de control de Cryptoblabes gnidiella. Phytoma. 298: 75-76. Anshelevich, L., Kehat, M., Dunkelblum, E. & Greenberg, S. 1993: Sex Pheromone Traps for Monitoring the Honeydew Moth, Cryptoblabes gnidiella: Effect of Pheromone Components, Pheromone Dose, Field Aging of Dispenser and Type of Trap on Male Captures. Phytoparasitica 21 (3): 189-198. Demirel, N. 2016: Seasonal flight patterns of the honeydew moth, Cryptoblabes gnidiella Millière (Lepidopetra: Pyralidae) in pomegranate orchards as observed using pheromone traps. Entomology and Applied Science Letters 3 (3):1-5. Öztürk, N. 2018: Creating a degree-day model of honeydew moth [Cryptoblabes gnidiella (Mill., 1867) (Lepidoptera:Pyralidae)] in pomegranate orchards. Türk. Entomol. Derg. 42 (1):53-62. Vidart, M.V., Mujica, M. V., Calvo, M. V, Duarte, F., Betancourt, C. M., Franco, J & Scatoni, I. 2013: Relationship between male moths of Cryptoblabes gnidiella (Millière) (Lepidoptera: Pyralidae) caught in sex pheromone traps and cumulative degree-days in vineyards in southern Uruguay. SpringerPlus 2(1): 1-8.

PheroFip 2019 62

Field effectiveness of pheromone-based mating disruption to control the vine mealybug, Planococcus ficus (Hemiptera: Pseudococcidae): results from Italy

Andrea Lucchi1*, Pompeo Suma2, Edith Ladurner3, Andrea Iodice3, Francesco Savino3, Renato Ricciardi1, Francesca Cosci1, Enrico Marchesini4, Giuseppe Conte1, Giovanni Benelli1

1Department of Agriculture, Food and Environment, University of Pisa, via del Borghetto 80, 56124 Pisa, Italy; 2Department of Agriculture, Food and Environment, University of Catania, via S. Sofia, 100, 95123 Catania, Italy; 3CBC (Europe) Srl, Biogard Division, via E. Majorana 2, 20834 Nova Milanese (MB), Italy; 4AGREA S.r.l. Centro Studi Via Garibaldi 5/16, 37057 San Giovanni Lupatoto (VR), Italy *[email protected]

Abstract Pheromone-based mating disruption (MD) has been tested on a wide range of insects of agricultural importance. The successful results obtained on a relevant number of pest species boosted its wide-scale employ, with a growing number of commercial products currently available to farmers. Nowadays, the vine mealybug (VMB), Planococcus ficus (Hemiptera: Pseudococcidae), is recognized as one of the main insect pests damaging vineyards. The harmfulness of VMB is due to phloem ingestion, resulting in degeneration of plants, decreased vigour, early defoliation, reduced bunch quality and decreased organoleptic characteristics of the obtained wine. This species produces a large amount of honeydew and wax, which serves as a substrate for sooty molt growing that reduces photosynthetic activity in leaves. Besides, VMB can transmit the grapevine leaf roll-associated virus and increases the risk of ochratoxin A contamination. A major challenge in vineyard IPM is reducing the amount of broad-spectrum insecticides currently used to manage insect pest populations, with special reference to moths, such as Lobesia botrana, and mealybugs. The chemical ecology of P. ficus has been extensively studied, with the identification of its sex pheromone in 2001 by Hinkens and co-authors, followed by the description of the site of pheromone emission in 2012. The effective employment of the pheromone main component, lavandulyl senecioate, for MD purposes was reported in 2006 and 2014 by Walton and colleagues and by Cocco and co-workers, respectively. In this scenario, little is still known about the optimization of MD programs against P. ficus. Herein, field research aimed at the successful management of VMB was recently carried out both on table grape and grapevine in various Italian regions. We focused on the potential impact of different amounts of pheromone (respectively 54, 72 and 90 g/ha) per hectare on VMB mating and thus VMB infestation levels. This was achieved by deploying per hectare respectively 300, 400 and 500 Isonet® PF reservoir dispensers, each dispenser containing 180 mg of racemic lavandulyl senecioate, and by assessing the effectiveness of the three different rates in reducing VMB infestation levels in comparison to control vineyards. Results pointed out that all three tested dispenser rates and thus amounts of pheromone per hectare were effective in reducing the percentage of VMB infested bunches as well as the number of VMB per bunch, with significant differences over the untreated control.

Key words: grapevine; Integrated Pest Management; mating disruption; mealybugs; sex pheromones

PheroFip 2019 63

Incorporating Mating Disruption into IPM Programs for Navel Orangeworm in California Almonds

David Haviland1, Jhalendra Rijal2

1University of California Cooperative Extension, Kern Co. 1031 South Mount Vernon, Bakersfield, CA 93307, USA, [email protected]; 2University of California Statewide Integrated Pest Management Program, 3800 Cornucopia Way, Suite A, Modesto, CA 95358, USA

Abstract Navel orangeworm (NOW), Amyelois transitella (Walker), is the most significant insect pest of more than 525,000 ha of almonds in California. Damage includes direct feeding on kernels and can predispose almonds to infection by Aspergillus sp. fungi which produce aflatoxins. Management of NOW requires a combination of integrated pest management (IPM) techniques to keep damage at levels that are commercially acceptable and allow for maximum economic returns to the grower. For more than a decade IPM programs focusing on sanitation and insecticides timed according to degree-day models did a good job keeping NOW at levels that are generally acceptable industrywide. However, over the past few years the tolerance for damage has decreased due to increased crop values, decreased tolerances for aflatoxins in some export markets, and regional changes in the agricultural ecosystem. Ten years ago California had approximately 300,000 ha of almond orchards that were typically surrounded by non-hosts for NOW, such as grapes or annual vegetable and field crops. Today, almonds are produced on over 525,000 ha, which when added to another 250,000 ha of pistachios and walnuts, means that many orchards are surrounded by other NOW hosts, increasing the need for coordinated areawide efforts against this pest. Mating disruption is a relatively new technique that provides an opportunity to improve NOW management within IPM programs. It is considered to be a low-risk, sustainable strategy, that can be used in complement with existing programs based on sanitation and traditional insecticides. Currently in California there are four different manufacturers with products that are commercially available to growers. These include two static-release dispenser systems using aerosol-based canisters from Suterra and Pacific Biocontrol, one variable-rate system using aerosol-based canisters where dispensers can be turned on and off remotely from Semios, and a static-release system using passive dispensers from Trécé. During 2017 we evaluated these four MD systems on 16-ha plots compared to an untreated check. These trials were funded by almond growers through the Almond Board of California. The MD systems were manufactured by four different companies and all released approximately the same amount of pheromone over the course of the season. The systems utilized active or passive dispersion of pheromone, different dispenser densities, and either static or variable rates of pheromone release. All systems were installed around April 1st onto 16-ha plots in three different orchards in Kern County, California. Each orchard had good winter sanitation and was sprayed once or twice with insecticides at hull split. In other words, we evaluated mating disruption as an added component to an existing IPM program, not as a replacement for sanitation or chemical control. All four MD systems caused greater than 90% reductions in the capture of males in pheromone traps from April through September. Egg captures during the same time period were reduced by 22% where MD was used. Across all orchards the average percentage of NOW-infested kernels was 2.28% for the no-MD checks compared to 1.13 to 1.33% for the

PheroFip 2019 64

four MD products. When all four MD products are averaged, MD reduced NOW damage in the varieties Nonpareil by 35%, Monterey by 51% and Fritz by 55%, with an overall reduction of 46%. Economic analyses were performed by calculating crop value using assumed per-ha yields of 1,704 kg of nonpareil at $5.50/kg and 1,704 kg of pollinizers at $4.95/kg base price, plus the addition of quality premiums according Blue Diamond’s 2017 Crop Quality Schedule. Where mating disruption was used, grower returns increased by an average of $246 per ha. This was comparable to the cost of implementing mating disruption. In other words, adding MD to existing management programs on a 16-ha scale pays for itself. In the process, levels of NOW damage for nuts arriving at the huller were cut in half and risks of aflatoxin were reduced. During 2017 we also conducted mating disruption demonstration trials to promote sustainable and environmentally-friendly pest management practices that were funded by the California Department of Pesticide Regulation. Six side-by-side comparison orchards were established in the Central Valley (Wasco, Maricopa, Lost Hills, Turlock, Escalon and Ballico). Each trial compared approximately 24 to 40 ha of conventionally-grown almonds to adjacent orchards of similar size where MD was used. Four of the orchards were rectangular, whereas the other two were used to see if MD would work in orchards oriented as triangles. Across all sites, MD reduced the number of male NOW caught in pheromone traps by 93% from April through September. The average percentage of NOW damage was 2.59% in conventional orchards compared to 1.14% where MD was included. This is a reduction of 55.7%. When only the four rectangular orchards are evaluated, moth captures were reduced by 96% and the average reduction in damage improved to 76.4%. When only considering the two triangular orchards, MD helped reduce male captures, but did not result in a reduction in damage at harvest. Evaluation of the economic benefits of MD showed that grower returns increased by an average of $268 per ha across all sites, and by $488 per ha in the four rectangular orchards. Either way you look at it, adding MD paid for itself. Additionally, at one location with relatively low NOW pressure (Wasco), the grower successfully omitted two insecticide sprays in the MD orchard and still had less damage than where the sprays were used. These economic benefits on approximate 40-ha orchards are even larger than the benefits shown previously on 16-ha plots, confirming a long-known fact that the efficacy of MD systems improves as the contiguous acreage under MD increases. All of our field work in 2017 suggests that incorporating MD for navel orangeworm in almonds in addition to an existing IPM program pays for itself when implemented on at least 16 acres, and generates a positive return on investment when implemented on at least 40 contiguous acres, all while reducing NOW damage and risk of aflatoxins. This statement assumes that the orchard is square to rectangular in shape. If the orchard has a high ratio of edges to middle (such as a triangle or a really long rectangle), growers who want to use MD should pair up with neighboring growers to make larger, more contiguous areas under MD to improve efficacy. Mating disruption is compatible with, and should be used in conjunction with (not instead of) other historically-proven management techniques: namely winter sanitation, insecticide sprays, and early harvest. Mating disruption is recognized as a sustainable, environmentally-friendly tool for controlling pests that goes hand in hand with industry-led efforts to produce food products for health-conscious consumers around the world.

Key words: integrated pest management, IPM, navel orangeworm, NOW, mating disruption

PheroFip 2019 65

On the shoulders of giants: mating disruption of Cydia latiferreana in hazelnuts

Betsey Miller1*, Daniel Dalton1, Vaughn Walton1

1Oregon State University, Department of Horticulture, 4017 Ag and Life Sciences Building, Corvallis, Oregon, 97331, USA *[email protected]

Abstract In 2016, global production of hazelnut, Corylus avellana was estimated at 397,000 metric tons (INC 2016). The United States holds a 3 to 4% share of global production (valued at $118 million), 99% of which is located in the state of Oregon (INC, 2016, ODA, 2018). The key pests of hazelnut in Oregon include filbertworm, Cydia lattiferreana (Lepidoptera: Torticidae) and filbert aphid, Myzocallis coryli. Filbertworm is a North American native and a close relative of the well-studied codling moth. It has numerous wild hosts, and, if left unmanaged, can infest up to 50% of nuts in a hazelnut orchard (Chambers et al., 2011). The economic threshold for filbertworm damage is less than 1%. The filbert aphid, once a major threat to the hazelnut industry, is now almost entirely controlled by the hymenopteran parasitoid Trioxy pallidus (Aphidiidae), which was imported from Europe in the 1980’s (Messing & AliNiazee, 1989). Hazelnut producers are well aware of the contribution of this parasitoid to their IPM programs and are otherwise concerned about overreliance on pyrethroid insecticides. In 2012, at the request of the industry and building upon decades of research on codling moth mating disruption, we began a five-year study to determine if mating disruption would be a suitable alternative to pyrethroid-based control of filbertworm. The sex pheromone of filbertworm (a mixture of E8, E10-12:OAc and E8, Z10-12:OAc), characterized in 1984, has been used to monitor filbertworm for decades. The current mating disruption product was developed by Shin Etsu Chemical Company, Ltd. (Tokyo, Japan) and is produced by Pacific Biocontrol Corporation (Litchfield Park, AZ, USA). Preliminary work by Hedstrom et al. (2014) determined the most appropriate dispenser (1-meter polyethylene tubes) and lowest effective rate for mating disruption (10 dispensers per acre). Our study was conducted on-farm in the McKenzie River Valley, Oregon, USA. Trials were designed to minimize edge effects by favoring plot size over number of replicates, with a minimum plot size of 12 hectares. Plots treated with mating disruption dispensers were compared with plots treated with pyrethroid insecticide applications. On all farms, growers were encouraged to spray insecticides, even in mating disruption plots, if counts of filbertworm in traps reached economic threshold. In 2012, increased filbertworm damage was observed at orchard perimeters, so in subsequent years, growers were encouraged to apply insecticides to orchard borders to protect against immigrating moths. In all plots, we monitored filbertworm flight with pheromone-baited traps and assessed percent infestation of nuts during harvest. In addition, we monitored filbert aphid populations and the percent parasitization by T. pallidus to assess differences in natural enemy activity between mating disrupted plots and those managed with insecticides. Successful disruption of mating, as indicated by trap shutdown, was inconsistent over the five-year period. In 2012 and 2013, trap shutdown was 87.5% and 88.3%, respectively. Over the next three years, it ranged from 41% to 62%. This breakdown in trap shutdown may be related to an increase in filbertworm density over the study period. From 2012 to 2015, the sum of filbertworm per trap in control traps increased incrementally each year from 1.6 in

PheroFip 2019 66

2012 to 12.6 in 2015; it decreased to 9.1 in 2016. In all five years, there was no significant difference in percent nut infestation between mating disruption plots and chemical control plots. Across all years, mean percent nut infestation was .47% in mating disrupted plots and .51% in chemical control plots. Despite the fact that mating disruption plots were sometimes treated with insecticides, the amount of insecticide applied to these plots was reduced by 45 to 75% compared to chemically managed plots. In all years, mean number of aphids per leaf was significantly greater in chemical control plots than in mating disruption plots and mean percent parasitism by T. pallidus was significantly greater in mating disruption plots in years 2012-2015. The economics of mating disruption (if border applications and periodic knockdown applications of insecticide are assumed) are comparable to that of chemical control. Coupled with the results from our study, this indicates potential for mating disruption as a suitable alternative to chemical control. Since the initiation of this study, the filbertworm pheromone dispenser has become a commercial product that is certified for use on organic farms. Industry wide adoption and success with this technology remains to be seen.

Key words: Mating disruption, hazelnut, filbertworm, biological control, parasitoid

Acknowledgements We thank Pacific Biocontrol, the Oregon Hazelnut Commission, Oregon State University’s Agricultural Research Foundation and the Eugene Water and Electric Board (Eugene, OR USA) for funding this research. We thank our collaborating growers, Jeff Newton, John Sullivan, Garry Rodakowski, Jim Goodpasture, Hugh Evonuk and Tom Evonuk, and Arlene Dietz for allowing us to work in their orchards.

References Chambers, U., Walton, V. M. & Mehlenbacher, S. A. 2011: Susceptibility of hazelnut cultivars to filbertworm, Cydia latiferreana. HortScience 46: 1377–1380. Hedstrom, C. S., Olsen, J., Walton, V. M. & Chambers, U. 2014: Pheromone mating disruption of filbertworm moth (Cydia latiferreana) in commercial hazelnut orchards. Acta Hortic. 1052: 253–262. https://doi.org/10.17660/ActaHortic.2014.1052.34. (INC) International Nut and Dried Fruit Council. 2016. Nuts and Dried Fruits Statistical Yearbook 2016-2017. Messing, R. H. & AliNiazee, M. T. 1989: Introduction and establishment of Trioxys pallidus [Hym.: Aphidiidae] in Oregon, U.S.A. for control of filbert aphid Myzocallis coryli [Hom.: Aphididae]. Entomophaga 34: 153-163. (ODA) Oregon Department of Agriculture. 2018: Oregon Agriculture: Facts and Figures. Oregon Department of Agriculture, Salem, Oregon. http://www.oregon.gov/ODA/shared/Documents/Publications/Administration/ORAgFact sFigures.pdf.

PheroFip 2019 67

Trials on Pheromone Mating disruption for reducing population of the Western Corn Rootworm, Diabrotica virgifera virgifera, (Coleoptera: Chrysomelidae)

Magda Rak Cizej1, Silvo Žveplan1, Iris Škerbot2

1 Slovenian Institute of Hop Research and Brewing, Plant Protection Department, C. Žalskega tabora 2, 3310 Žalec, Slovenia; [email protected]; 2Agriculture and Forestry Chamber of Slovenia, Agricultural and Forestry Institute Celje Trnoveljska cesta 1, 3000 Celje, Slovenia

Abstract Preliminary results for reducing population of the Western corn rootworm beetle, Diabrotoca virgifera virgifera (Dvv), with sex pheromone in maize fields in Slovenia are reported. Research activities carried out in years 2015-2018 and where focused on monitoring and mating disruption of adults Dvv. Spermatophores dissected from the females and maize root injury were checked. Sex pheromone was applied with tractor sprayer when appeared the first male adult Dvv on pheromone traps. According to our preliminary results the mating disruption can be good control systems for Dvv according with the guidelines of Integrated Pest Management.

Introduction The Western Corn Rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) is an important pest of maize (Zea mays), present in North America and also in Europe, whose soil-inhabiting larvae can seriously damage roots of maize and lead to yield losses. Dvv is present in Slovenia from 2003 and populations growing up every year because maize is important host plant which grown on 40% fields every year. The studies present results for using sex pheromone-mating disruption on two different locations for reducing population of the Dvv.

Material and methods Monitoring: On two locations in Slovenia, Draža vas by Slovenske Konjice (2015) and Savci by Mala Nedelja (2017) carried out monitoring of female Dvv with pheromone PAL trap Csalomon® on maize fields from June to September. We counted number of males adult in pheromone traps each week. The pheromone replaced every month. Mating disruption (MD): The MD trials carried out on location Draža vas by Slovenske Konjice (2015) Savci by Mala Nedelja (2017) for reducing population Dvv on maize. On each location have used two treatments; untreated and treated plot where used synthetic sex pheromone 8-methyl-2-decanol propanoate which applied on volcanic stone zeolite (product CornProtect, Lithos Natural, Austria) in dose of 4 kg/ha with 300 l water/ha. Foliar application made by tractor sprayer when maize plants were in average 1.2 m height. On location Draža vas by Slovenske Konjice in year 2015 evaluated biological efficacy of MD where collected target pest Dvv beetles every seven days from first flight activity (2 July 2015) to end of flight activity (10 September 2015). In each sampling, minimum 100 females and 100 males adults of Dvv randomly collected in the middle of the untreated plot and treated experimental plot and add give in bottle with 70% alcohol. In the laboratory, the male and females were counted and rate of the male and female adults were determined. Then the spermatophores were dissected from female under stereomicroscope and spermium content of

PheroFip 2019 68

the spermatophores were examined and numbers of sterile females were counted in the treatments in each of collecting dates. Root injury /damage assessment: In 2016 (Draža vas by Slovenske Konjice) and 2018 (Savci by Mala Nedelja) collected 60 roots (15 roots *4 replications) on each treatment. Root damage on maize assessed by node-injury scale.

Results and discussion In the present experiment, which was carried out on the fields in of Draža vas near Slovenske Konjice, Slovenia (app. 10 ha), in years 2015, and Savci by Mala Nedelja in year 2017 where population of Dvv adults were very high, used mating disruption technique with the sex pheromone 8-metil- 2 decanolpropanoat was examined. In 2015 on location Draža vas near Slovenske Konjice we detected reduced mating of males (females were 46% sterile). The field experiments demonstrated that after MD females Dvv layed fewer eggs in soil and we observed in average up to 23% less damage on the roots of maize in year after MD using. In the same year of MD using less damage on maize silks observed (in average 7 %). Population of Dvv are very high in Slovenian maize fields therefore the use of continuous rotation is highly recommended for managing Dvv populations. At the same time the combination of safe and environmental friendly crop management practice with the guidelines of Integrated Pest Management using biological control agents (entomopathogenic nematodes) or biotechnology-MD adapted to specific maize production systems in Slovenia is highly recommended.

Key words: Diabrotica virgifera virgifera, pheromones, monitoring, mating disruption, spermatophora dissected

PheroFip 2019 69

Predicting pine processionary moth outbreaks with pheromone trapping

H. Jactel1, I. van Halder1, E. Kersaudy2, T. Belouard2

1INRA, umr Biogeco, Laboratory of Forest Entomology, Cestas, France; 2Department of Forest Health, Bordeaux, France

Abstract The pine processionary moth (PPM, Thaumetopoea pityocampa) is considered the most severe insect defoliator of pine species in southern Europe and North Africa, with large impacts on forest vitality and productivity. PPM exhibits periodic outbreaks, with a return period of seven to nine years. However, the amplitude of outbreaks is not predictable. Following the identification and synthesis of the sex pheromone in the 1980’s we proposed a standardized pheromone trapping system for the monitoring of PPM infestations, based on plate sticky traps baited with 0.2mg of pityolure. The aim of the current study was the optimization of pheromone trapping networks for the prediction of PPM damage, based on correlation between male captures and level of infestation in the following generation. We set up a long-term experiment with a systematic network of 50 plots in a large maritime pine plantation forest of one million ha (hereafter regional level) in southwestern France. Sampled plots were aligned on a 16 x 16km grid and surveyed for the percentage of PPM attacked trees from 2010 to 2016. During the same period, four sticky traps were installed in each plot and assessed four times during the flight period for number of caught males. We estimated correlations between sum trap captures per year and infestations in the following generation. While correlations at the plot level were not significant for a given year, correlations between mean trap capture and mean percentage of attacked trees per year were positive and highly significant at the regional level for the seven years period. Using only two traps out of four and two assessments period out of four to estimate sum trap capture did not reduce the level of correlation. Applying random resampling procedure among plots, we further demonstrated that temporal correlations remained positive and highly significant with only eight sampled plots and associated two traps in the region. Our findings suggest that a low density network of pheromone baited traps, assessed for a short period of time at the beginning of flight period, could provide a good estimate of PPM infestations in the following generation, and thus of defoliation damage, at the regional level. This paves the way for a simple and practical method of early warning for the initiation of preventive control measures against the pest.

Key words: pheromone trap, systematic network, prediction, outbreak, defoliator

PheroFip 2019 70

The eucalyptus weevil Gonipterus platensis (Coleoptera, Curculionidae): new control perspectives based on semiochemicals

Sofia Branco1, Eduardo Mateus1, Stefan Schütz2, Maria Rosa Paiva1

1CENSE- Center for Environmental and Sustainability Research, DCEA, Faculdade de Ciências e Tecnologia (FCT), Universidade Nova de Lisboa (UNL),Caparica Portugal; 2Department of Forest Zoology and Forest Conservation, Buesgen-Institute Göttingen University, Göttingen, Germany

Abstracts The eucalyptus weevil Gonipterus platensis, is a defoliator originating from Oceania, that has spread and invaded large areas where eucalyptus are planted for commercial purposes in Africa, Europe and America (Reis et al. 2012). Attacks of this insect to eucalyptus in several regions often reach values that exceed the economic level of damage, as both larva and adults feed on eucalyptus leaves and young shoots, leading to reduced tree growth and tree mortality. In some climatic regions, the beetle can be controlled by a hymenopteran of the genus Anaphes, which is an efficient egg parasitoid (Tooke, 1953). However, in colder and wetter regions, the efficiency of the parasitoid decreases, so that control of the weevil is not achieved (Reis et al. 2012). In Portugal, G. platensis reaches pest status in approximately 18.5% of the 812 000 hectares planted with eucalyptus, and has caused a wood loss of about 648.M euros over the last 20 years (Valente et al. 2018). The use of semiochemicals for integrated pest management (IPM) of Gonipterus species is still largely unexplored. In this study we further present a prospect for biotechnological control based on compounds released by its host Eucalyptus globulus that might mediate host selection behaviour in this weevil species. In this study E. globulus volatile organic compounds were collected by headspace solid phase microextraction, monolithic material sorption extraction and simultaneous distillation- extraction. Gas chromatography - mass spectrometry/ electroantennographic detection (GC- MS/EAD) was used to determine which compounds triggered antennal responses in G. platensis. Further chemical analysis of the extracted compounds was conducted using gas chromatography and mass spectrometry and comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry. G. platensis antennal response was elicited by 51 host plant volatiles, and was confirmed for 33 of these compounds using commercial standards. The behavioural response of G. platensis to individual compounds was assessed in a two-arm olfactometer with ten parallel walking chambers, coupled to video tracking and data analysis software. Weevils of known age, sex and mating status were tested. We identified eight compounds with kairomonal activity and two compounds with allomonal property. The behavioural responses were related to the sex and mated condition of the tested insects. Three compounds attracted virgin females or both sexes: camphene, (+)-α-pinene and 2-phenylethanol, and are potential candidates for application in integrated pest management approaches. Furthermore, the two allomones identified may have potential use for developing push-pull strategies (Branco et al. 2018). In short, sustainable management of G. platensis can only be achieved through the adoption of an integrated approach, resorting to an array of techniques: i) Population monitoring based on sampling, coupled with teledetection to assess defoliation rates and inform decision making on treatment applications; ii) Genetic selection of eucalyptus clones showing reduced susceptibility to the weevil; iii) Release/augmentation of biological control agents, in combination with biotechnological strategies; iv) Biotechnological control, based

PheroFip 2019 71

on the use of semiochemicals - both volatile organic compounds (VOCs) released by the host trees as well as pheromones emitted by the weevils, having attractant, deterrent or repellant effects on conspecifics and/or natural enemies. In general, research conducted to develop the proposed tools yielded promising results and is in progress. The strategies envisaged to be implemented in eucalyptus plantations include push–pull techniques, use of trap crops, attraction of natural enemies, weevil repellency and/or oviposition deterrence.

Key words: kairomones, allomones, Eucalyptus globulus, Gonipterus platensis

Acknowledgements Sofia Branco received a Ph.D. scholarship from Fundação para a Ciência e Tecnologia (FCT)—SFRH/BD/84412/2012. This work has been supported by CENSE (Center for Environmental and Sustainability Research) which is financed by national funds from FCT/MEC (UID/AMB/04085/2013).

References Branco, S., Mateus, E. P., da Silva, M. D. G., Mendes, D., Rocha, S., Mendel, Z., Schütz, S. & Paiva, M. R. 2018: Electrophysiological and behavioural responses of the Eucalyptus weevil, Gonipterus platensis, to host plant volatiles. Journal of Pest Science 1-15. Reis, A. R., Ferreira, L., Tomé, M., Araujo, C. & Branco, M. 2012: Efficiency of biological control of Gonipterus platensis (Coleoptera: Curculionidae) by Anaphes nitens (Hymenoptera: Mymaridae) in cold areas of the Iberian Peninsula: implications for defoliation and wood production in Eucalyptus globulus. Forest Ecology and management 270: 216-222. Valente, C., Gonçalves, C. I., Monteiro, F., Gaspar, J., Silva, M., Sottomayor, M., Paiva, M. R. & Branco, M. 2018: Economic Outcome of Classical Biological Control: A Case Study on the Eucalyptus Snout Beetle, Gonipterus platensis, and the Parasitoid Anaphes nitens. Ecological Economics 149: 40-47. Tooke, F.G.C. 1953: The Eucalyptus Snout. beetle, Gonipterus scutellatus Gyll. A Study of its Ecology and Control by biological Means. E. M. of the Department & U.S.A. of Agriculture, eds. Entomology Memoirs Department of Agriculture Union of South Africa 3:1–282.

PheroFip 2019 72

SESSION 5

SIT AND BIOLOGICAL CONTROL OF

FRUIT CROP PESTS

CHAIRPERSON: KERSTIN KRÜGER

PheroFip 2019 73

Area-wide Management of Fruit Flies Using the Sterile Insect Technique

Rui Pereira1*, Walther Enkerlin1, Carlos Cáceres1, Daguang Lu1, Marc J.B. Vreysen1

1Insect Pest Control Section, Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture, Vienna, Austria *[email protected]

Abstract Traditional control of fruit flies is mostly based on the uncoordinated application of insecticides using an orchard-by-orchard approach. This strategy is not effective or sustainable in view of fruit fly movement and is also damaging for the environment. The area- wide (AW) approach has its roots in the management of vector-borne diseases and locust pests, and has increasingly been used for the management of fruit flies. A fruit fly area-wide integrated pest management (AW-IPM) strategy targets the entire pest population which requires treating all habitats to avoid leaving significant infestations from where migrant insects can re-establish in areas of concern. Insect movement, occurring sometimes over long distances, is generally underestimated. Consequently, most conventional pest management is an uncoordinated local action against only segments of a pest population, resulting very often in unsustainable control. On the other hand, an AW-IPM approach involves a preventive rather than a reactive strategy, whereby all individuals of the pest population are targeted in time and space, resulting in more cost-effective and sustainable pest management (Vreysen et al., 2007). The AW management requires coordination among all stakeholders, long-term commitment, usually multiyear planning, and a centralized organization dedicated exclusively to its implementation. The successful implementation of these programmes also requires adoption of a “phased conditional approach” where progression to the next phase is made conditional upon having finalised all or most of the required activities in the previous phase. This entails collection of necessary baseline data including species present, temporal and spatial dynamics of the target populations over several years, host status, host sequence. Other relevant information depending on the objective of the programme is, the fruit market targeted, the ecological characteristics of the production area and cost-effective control tactics available to be selected for integration in the programme. Among the control tactics available, the sterile insect technique (SIT) can contribute to the AW-IPM approach in situations where the number of key pests is low, and when integrated with other control methods. Among the methods available for integration, depending on the characteristics of the infested area are sanitation, bait sprays, mass-trapping, and male annihilation technique (MAT) (Dyck et al., 2005). The SIT is a method of pest control involving area-wide inundative releases of sterile insects to reduce reproduction in a field target population of the same species in an area. It is one of the most environment-friendly methods available, as no toxic material is introduced in the target area, and the released insects cannot get established, as sometimes happens with some other biological control methods. The AW-IPM with an SIT component against fruit flies can be implemented using four different strategies, i.e. suppression, containment, eradication, and prevention. With the markets growing that require lower pesticide residues, the SIT has been used more and more for fruit fly suppression replacing conventional control. This strategy, can also be used to maintain an area of low fruit fly prevalence that can be complemented with a post-

PheroFip 2019 74

harvest phytosanitary treatment, i.e. an additional independent risk mitigation measure, as part of a systems approach to reach “pest free markets” (FAO, 2012). This is the current strategy being used against the Mediterranean fruit fly (or medfly) Ceratitis capitata, Wied., by the citrus producers in Valencia, Spain to get access to the lucrative USA markets. Fruit fly containment is used mainly to limit the progress of the fruit flies to contiguous regions that are free of the pest. An example of this strategy is the release of millions of sterile medfly males along the Mexico-Guatemala border that has maintained Mexico free of this pest for the past four decades. Initially, eradication was the main goal of SIT projects, this days eradication is mainly used for special situations where it is technically and economically justifiable. Among those situations, the incipient introductions of invasive species in new areas has received recently special attention. For example, a medfly outbreak in the Dominican Republic triggered an instant ban to fruit and vegetable exports. The Minister of Agriculture with the support of FAO, IAEA, USDA and other stakeholders reacted immediately with the implementation of an eradication effort using an AW-IPM approach integrating the SIT. The successful sustainable removal of the medfly from the Dominican Republic reopened the fruit and vegetable export markets to the USA valued at that time in US$ 40 million. In these situations, the detection and rapid response to the outbreak is crucial to the successful eradication, irrespective of the control tactics used (FAO, 2016; FAO/IAEA, 2018). Although the strategy of prevention has been used less extensively, this approach (Mediterranean Fruit Fly Preventive Release Program (PRP) in California and Florida) has in the last decades prevented the establishment of this pest in the continental USA (Dowell et al. 2000). Finally, the challenges that AW-IPM face and the integration of SIT with other control methods will be discussed.

Key Words: Area-wide Integrated Pest Management (AW-IPM), Phased Conditional Approach, Suppression, Containment, Eradication, Prevention

References Dyck V. A., Robinson, A. S. & Hendrichs, J. 2005: Sterile Insect Technique Principles and Practice in Area-Wide Integrated Pest Management, The Netherlands. 787 pp. FAO 2012: Systems Approach for Pest Risk Management of Fruit Flies. ISPM No. 35, International Plant Protection Convention (IPPC). FAO, Rome, Italy. FAO 2016: Establishment of Pest Free Areas for Fruit Flies (Tephritidae). ISPM No. 26, International Plant Protection Convention (IPPC). FAO, Rome, Italy. FAO/IAEA 2018: Trapping guidelines for area-wide fruit fly programmes, Second edition, by Enkerlin, W. R. & Reyes-Flores, J. (eds). Rome, Italy. 65 pp. Vreysen, M. J. B., Robinson, A. S. & Hendrichs, J. 2007: Area- Wide Control of Insect pests from Research to Field Implementation. Springer, Dordrecht, The Netherlands. 789 pp.

PheroFip 2019 75

Back to the future: SIT in synergism with biological control

Massimo Cristofaro1, Claudio Ioriatti2, Gianfranco Anfora2, Valerio Mazzoni2, Gino Angeli2, Silvia Arnone3, Gerardo Roselli4, Kim Hoelmer5, Max Suckling6

1ENEA & BBCA, Rome, Italy; 2FEM, San Michele all´Adige (TN), Italy; 3ENEA Casaccia, Rome, Italy; 4BBCA-onlus, Rome, Italy; 5USDA ARS, Newark (DL) USA; 6Plant & Food research Institute, Auckland, New Zealand

Abstract Medfly (Ceratitis capitata), spotted wing drosophila (Drosophila suzukii) and brown marmorated stink bug (Halyomorpha halys) are invasive pest species in Europe. All are multivoltine and extremely polyphagous. For this reason, they are considered among the most severe pests in agriculture. The development of Sterile Insect Technique (SIT) programs, in combination with both classical and augmentative biological control, is in progress to verify the possibility to suppress the populations of these three pest species under a sustainable economic threshold, in two regions of North and Central Italy. Special emphasis is given to enhance the synergism between least toxic agricultural pest management, georeferenced monitoring of the territorial distribution of the pests and their natural enemies, and SIT applications both in area-wide and confined field conditions.

PheroFip 2019 76

Optimizing the sterile insect technique for management of codling moth

Larry Gut1, Rob Curtiss1, Chris Adams1

1Department of Entomology, Michigan State University, East Lansing MI USA 48824

Abstract Codling moth (CM), Cydia pomonella (L.), is a primary pest of apple, pear, and walnut worldwide. CM directly feeds on fruit and losses in revenue from CM injury and costs of controlling this pest can be significant. There are several options for controlling CM, including insecticides, mating disruption, and virus. However, the efficacy of these tactics as a stand-alone control has diminished in the past five years. CM is once again a primary pest due to loss of effective insecticides and increasing pest pressure overwhelming mating disruption and other biopesticides. New tactics for controlling this pest are sorely needed. For several insect pests, including CM, the sterile insect technique (SIT) has proved effective as a means of greatly reducing population densities over a wide area (Klassen and Curtis 2005). The approach entails sterilization of reared insects that are subsequently released in large numbers to compete with wild individuals to reduce or even eliminate fertile matings and thus, production of offspring (Lance and McInnis 2005). An areawide program to suppress CM using a combination of SIT, pheromone-based mating disruption and judicious use of insecticides has been conducted since 1999 in the pome fruit producing areas of the south-central part of British Columbia. The approach has reduced insecticide sprays and CM damage across the region. The British Columbia SIT program has initiated a program whereby growers outside of Canada would have access to sterile moths for managing CM. The intent is that sterile moths could be applied to individual orchards much like an insecticide spray. In order to make this a cost-effective alternative for managing CM, research is needed to optimize the methods and timing of release of sterile adults, as well as the impact of farm practices on their effectiveness. In the Canadian SIT program, moths are released uniformly throughout each apple orchard using a modified 4-wheeler. The approach is time consuming and costly. Our previous research on CM movement in apple orchards revealed that released males spread quickly over a fairly large area, dispersing more than half a hectare in a single evening. Thus, we conducted several experiments to determine the impact of release method on the distribution of moths following their release. Over the course of the summer, more than 1.6 million sterile CM were released in carrying out a dozen experiments. We compared release of SIR moths from an unmanned aerial vehicle (UAV or drone) or from the ground and compared releasing moths by hand from a single central site vs uniform release from multiple locations within the orchard. Experiments were conducted in 10-acre plots parceled out of the center of large orchards to provide a buffer between treatments. The distribution of released moths was assessed by recapturing them in CM/DA/AA baited Delta traps deployed in a grid within each plot. For each treatment, moths were marked with a unique fluorescent dye and released at a rate of 800/acre (400 males, 400 females). A single central or spread release by hand provided very similar and uniform distributions of SIR moths. The spread release of moths from a drone also provided a uniform distribution. However, the distribution of moths following a single release from the drone was not uniform. Moth catch was skewed in a downwind direction. Moths are currently released from the drone flying at a height of 35m. Our plans for 2019 are to determine if releasing moths at a lower altitude will mitigate the effect of wind and improve distribution.

PheroFip 2019 77

In a companion experiment, we compared moth distribution following hand releases from a single central location, four corners of the plot and spread evenly across 20 locations. 8000 sterile moths were release in each plot, and the experiment was replicated nine times. The three approaches to hand releasing sterile moths produced similar and uniform patterns of recapture. Our evaluation of the options for releasing moths for the purposes of SIR conducted over the past two years reveal that releasing moths from only a few locations is just as effective as evenly distributing them across the entire block. Moths exhibit the ability to quickly disperse throughout the orchard once they have been released from only a limited number of locations in the orchard. This outcome is good news for the economics of CM SIT. While CM adults are only searching for mates for a few hours at dusk and into the early evening, sterile moths may be released anytime during the day. Does the time of release impact the performance and distribution of SIR moths? To answer this question, we compared moth survival and distribution following their release at times ranging from 12 PM to 9 PM (noon to dusk). SIR moths were released at a rate of 800/acre from a central location at 3-hour intervals beginning at noon. Moths were recaptured in 20 CM/DA/AA baited Delta traps placed in a 10-acre grid. Trials conducted in both the spring and summer indicated that recaptures rate were substantially higher when moths were released earlier in the day rather than at dusk or nighttime. Releases just prior to or during the dusk activity period appears to be suboptimal for moth survival and distribution. Earlier releases may allow the SIR moths to acclimate and be ready to search for mates during the first night after release. In contrast, moths released late in the day may be inactive for a while and mortality may be high. It is becoming more commonplace for growers to use coverings, such as netting, to protect their high value fruit crops. Thus, in a paired experiment we examined the effect of two heights of netting on the distribution of released moths. SIR moths were deployed in 4 ha trellised apple orchards covered with 6.6 meter or 3.3 meter high netting. In both orchards, 8000 fluorescent-dyed sterile codling moths were centrally released, and moth distribution was assessed by recapturing them in CM/DA/AA baited delta traps placed in an evenly spaced grid approximately 60’ apart. In the orchard covered by the taller netting there was at least 2 meters of open space above the tops of the apple trees. Under these conditions, moth movement did not appear to be affected by the netting, leading to a fairly uniform distribution. In contrast, the 3.3 m high netting was in contact with the upper canopy of the orchard. This situation appeared to interfere with moth movement. A substantial portion of the released moths were recaptured near the central point of release. Trials using a similar experimental design examined the effect of orchard architecture (trellis vs free-standing) and mating disruption treatment (hand-applied or aerosol emitters) on moth distribution. Surprisingly, SIR moths freely moved through the orchard despite the possibiltiy of their movement being impeded by a dense trellised planting. Similarly, moths distributed themselves uniformly even in the presence of pheromone treatments.

Key words: sterile insect, SIT, SIR, Cydia pomonella, integrated control

PheroFip 2019 78

Hoverflies as biological control agents of the rosy apple aphid in Mediterranean areas

Neus Rodríguez-Gasol1, Jesús Avilla2, Simó Alegre1, Georgina Alins1

1IRTA Fruitcentre, PCiTAL, Park of Gardeny, Fruitcentre Building, Lleida 25003, Spain 2Department of Crop and Forest Science, Agrotecnio, University of Lleida, Avda. Alcalde Rovira Roure 191, Lleida 25199, Spain [email protected], [email protected], [email protected], [email protected]

Abstract Hoverflies (Diptera: Syrphidae) are an important group of beneficial arthropods due to its double function as providers of ecosystem services like pollination and biological control of some pests. Adult hoverflies need to feed on pollen and nectar for survival and sex maturation, respectively (Gilbert, 1981). This dependence on flower resources has been researched and used in habitat manipulation schemes to enhance agroecosystem services in farmlands (Landis et al. 2000; Wratten et al. 2012). However, most of these studies have been conducted in colder regions and the plant species and their attractiveness may differ in warmer and drier areas such as the Mediterranean. It is precisely in these areas where aphids like the rosy apple aphid (Dysaphis plantaginea) are major pests of apple (Malus domestica Borkh.) orchards (Rousselin et al. 2017). In order to improve habitat management measures for aphid biological control, it is necessary to have a better knowledge of the aphidophagous hoverflies present in the area and the flower resources they use. The aim of this work was to improve biological control of the rosy apple aphid. For this reason we described the hoverfly species present in the fruit tree growing area of Lleida (Catalonia, NE-Spain), we identified the most attractive spontaneous flower species to hoverflies and we identified which hoverfly species predate the rosy apple aphid.

Material and methods Hoverflies naturally present in the area and their preferred spontaneous flora During April and May of 2016 and 2017 adult hoverflies visiting the spontaneous flora in different semi-natural habitats from the fruit tree growing area of Lleida were captured fortnightly by sweep netting in order to identify the hoverflies present in the area. The captured individuals were stored in glass pots with etil-acetate and the flower species where they were captured was noted. Once at the laboratory the individuals were identified to genera. Hoverflies found in the rosy apple aphid colonies In order to identify the hoverflies present in the rosy apple aphid colonies eight organic apple orchards were sampled fortnightly from April to May of 2018. In each orchard, 12 infested shoots were collected, kept in plastic glasses with fabric lids and taken back to the laboratory (at 20ºC temperature and 12:12 (L:D) h). Shoots were left for approximately two weeks to allow the immature stages of the hoverflies to develop. After, the individuals were identified to genera.

Results A total number of 65 individuals were captured on the spontaneous flora during the sampling period. These belonged to six different genera: Sphaerophoria (38.5%), Episyrphus (23.1%),

PheroFip 2019 79

Eupeodes (15.4%) Melanostoma (12.3%), Paragus (6.2%), and Xanthogramma (4.6%). The captured hoverflies visited 25 different plant species, belonging to 11 families. The most visited families were Compositae (52.3%) and Cruciferae (20.0%), being Anacyclus clavatus (29.2%) and Diplotaxis erucoides (9.2%) the most visited plants. Regarding the hoverflies present in the rosy apple aphid colonies, we found 67 adults that belonged to three different genera: Episyrphus (59.7%), Eupeodes (20.9%), and Sphaerophoria (19.4%).

Discussion This study shows that the diversity of hoverflies naturally occurring in semi-natural Mediterranean areas has potential to control rosy apple aphid populations since the genera found in the colonies were the most abundant feeding on spontaneous flora. These findings highlight the importance of spontaneous flora on hoverflies. However, few floral resources are present in orchard groundcover because farmers usually mown or shredded it. For this reason, efforts should be devoted to juggle weed control and floral promotion in order to increase biological control and, therefore environmental crop sustainability.

Key words: hoverfly, Syrphidae, Dysaphis plantaginea, ecosystem services, pests control, apple

Acknowledgements This research was funded by the Spanish projects RTA 2013-00039-C03-00 and AGL2016- 77373-C2, and the CERCA Programme/Generalitat de Catalunya.

References Gilbert, F.S. 1981: Foraging ecology of hoverflies - Morphology of the mouthparts in relation to feeding on nectar and pollen in some common urban species. Ecol. Entomol. 6: 245- 262. Landis, D. A., Wratten, S. D. & Gurr, G. M. 2000: Habitat management to conserve natural enemies of arthropod pests in agriculture. Annu Rev Entomol. 45: 175-201. Rousselin, A., Bevacqua, D., Sauge, M. H., Lescourret, F., Mody, K. & Jordan, M. O. 2017: Harnessing the aphid life cycle to reduce insecticide reliance in apple and peach orchards. A review. Agron Sustain Dev. 37: 13. Wratten, S. D., Gillespie, M., Decourtye, A., Mader, E. & Desneux, N. 2012: Pollinator habitat enhancement: Benefits to other ecosystem services. Agric Ecosyst Environ. 159: 112-122.

PheroFip 2019 80

Intraguild predation among natural enemies of Myzus persicae in peach trees in the NE of Spain

Yahana Aparicio1, Rosa Gabarra1, Judit Arnó1

1IRTA, Ctra. Cabrils km 2, 08348 Cabrils (Barcelona), Spain [email protected]; [email protected]; [email protected]

Abstract Spain is one of the main European peach producers with almost the 30% of the total production. Within Spain, in 2017, 30% of the total production was harvested in Catalonia, northeast of Spain. (FAOSTAD, 2016; MAPAMA, 2017). The green peach aphid, Myzus persicae Sulzer (Hemiptera: Aphididae) is the most important aphid species affecting peach trees. Nowadays, its control is mainly relying in insecticides sprays, but resistance to a wide range of them has been recorded (Slater et al. 2012). This, together with the social concern for health protection and an environmentally friendly agriculture, demands to develop alternative tools that enables to reduce the use of pesticides. Conservation biological control (CBC) may play an important role in the management of this pest since aphids have a wide range of natural enemies (NE). The inclusion of floral resources near crops may enhance NE populations by providing them with food resources and refuge (Brennan, 2016). Regular field samplings conducted during 2015 and 2016 in this area, showed that the main NE present were parasitoid wasps and the predators Orius spp., gall midges and hoverflies (Aparicio et al. 2015). The possibilities of CBC success may be affected by the possible interactions between the different NE present. These interactions are usually studied from the parasitoid point of view, because predators can attack parasitoids, but parasitoids are not able to attack predators (Colfer & Rosenheim, 2001). In this work we studied in laboratory trials the potential of the predators in aphid control and the interaction between them and parasitoid wasps. To do that, a female of the bug Orius majusculus (Hemiptera: Anthocoridae), a larvae of the gall midge Aphidoletes aphidimyza (Diptera: Cecidomyiidae) and a larvae of the hoverfly Episyrphus balteatus (Diptera: Syrphidae) were caged in a microcosm containing an infested sweet pepper plant during three days. After this time the number of alive aphids were counted. Our results showed that the hoverfly larvae was the most effective predator, decreasing the aphid population by 93%. The other predators consumed a lower amount of aphids. Thus, the gall midge reduced the aphid population by 20% and the O. majusculus by 10%. It has long been known that the hoverfly larvae can feed on parasitized aphids (Meyhöfer & Klug, 2002). However, there is few information regarding the interaction between the other predators and aphid parasitoids. Thus, we carried out two experiments to study the food preference of O. majusculus and A. aphidimyza between healthy and parasitized aphids. We performed a choice trial with a final count at 24 h and a “cafeteria” test in which several counts were performed during 48 h to see the order in which the prey was eaten by the predators. Our results regarding A. aphidimyza showed that the predator was able to feed on both although consumed more healthy aphids and consumed them earlier than the parasitized ones. Same trend was observed in the experiments done with O. majusculus. The results obtained in our study suggest that the main NE present in peach orchards in the study area could be valuable potential biological control agents of the green peach aphid and that interactions among A. aphidimyza and O. majusculus and the parasitoid wasps might not be detrimental to the biological control of the pest. The provision of food resources, such

PheroFip 2019 81

as flower nectar close to the orchards may be a useful strategy to enhance the abundance and early presence of parasitoid wasps (Arnó et al. 2012) and Orius spp. (Pumariño et al. 2012). Also A. aphidimyza improve their fitness when feeding on nectar. However, the morphology of predator adult prevents it from reaching the nectar of some flowers (Aparicio et al. 2018).

Key words: Predators, Aphidiinae, Green peach Aphid, association

Acknowledgements This research was supported by the Spanish Ministry of Economy and Competitiveness (Projects AGL2013-49164-C2-2-R and AGL2016-77373-C2-1-R) and the CERCA Programme/Generalitat de Catalunya.

References Aparicio, Y., Gabarra, R., Riudavets, J., Artigues, M., Rodríguez, N., Alins, G., Vilas, M. & Arnó, J. 2015: Enemigos naturales asociados a Myzus persicae (Sulzer) en cultivo de melocotón. IX Congreso Nacional de Entomología Aplicada / XV Jornadas Científicas de la SEEA. Valencia, Spain. Aparicio, Y., Gabarra, R. & Arnó, J. 2018: Attraction of Aphidius ervi (Hymenoptera: Braconidae) and Aphidoletes aphidimyza (Diptera: Cecidomyiidae) to sweet alyssum and assessment of plant resources effects on their fitness. J. Econ. Entomol. 111: 533-541. Arnó, J., Gabarra, R. & Alomar, O. 2012: Hymenoptera abundance on candidate plants for conservation biological control. IOBC-WPRS Bull. 75: 13-16. Blackman, R. L. & Eastop, V. F. 2000: Aphids on the World's crops. John Wiley & Sons, Chichester, UK. Bonsall, M. B. & Holt, R. D. 2003: The effects of enrichment on the dynamics of apparent competitive interactions in stage-structured systems. Amer. Naturalist 162: 780–795. Brennan, E. 2016: Agronomy of strip intercropping broccoli with alyssum for biological control of aphids. Biol. Control 97: 109-119. Colfer, R.G. & Rosenheim, J.A. 2001: Predation on immature parasitoids and its impact on aphid suppression. Oecol. 126: 292–304 FAOSTAT. Food and Agriculture Organization of the United Nations. 2016: Crops. (May 28, 2018). Last accessed: August, 24, 2018. URL: http://www.fao.org/faostat/en/#data/QC. MAPAMA (Ministerio de Agricultura, Pesca y Alimentación). (2017). Encuesta sobre Superficies y Rendimientos de Cultivos. Retrieved from: https://www.mapama.gob.es/es/estadistica/temas/estadisticasagrarias/boletin2017sm_tcm 30-455983.pdf. Last accessed August, 24, 2018 Meyhöfer, R. & Klug, T. 2002: Intraguild predation on the aphid parasitoid Lysiphlebus fabarum (Marshall) (Hymenoptera: Aphidiidae): Mortality risks and behavioral decisions made under the threats of predation. Biol. Control 25: 239-248. Pumariño, L. & Alomar, O. 2012: The role of omnivory in the conservation of predators: Orius majusculus (Heteroptera: Anthocoridae) on sweet alyssum. Biol. Control 62: 24- 28. Slater, R., Paul, V. L., Andrews, M., Garbay, M. & Camblin, P. 2012: Identifying the presence of neonicotinoid resistant peach-potato aphid (Myzus persicae) in the peach- growing regions of southern France and northern Spain. Pest Manag. Sci. 68: 634-638.

PheroFip 2019 82

Developing a biological control strategy for Cacopsylla spp. with novel entomopathogenic fungus Pandora sp.

Louisa M. Görg1,3*, Annette H. Jensen2, Jørgen Eilenberg2, Jürgen Gross1,3

1Julius Kühn-Institut, Federal Research Centre for Cultivated Plants, Institute for Plant Protection in Fruit Crops and Viticulture, Laboratory of Applied Chemical Ecology, Schwabenheimer Str. 101, Dossenheim, Germany; 2University of Copenhagen, Department of Plant and Environmental Sciences, Copenhagen, Denmark; 3Technical University of Darmstadt, Chemical Plant Ecology, Schnittspahnstr. 10, Darmstadt, Germany *[email protected]

Abstract A novel fungus Pandora sp. (Entomophthorales: Entomophthoraceae) was isolated from pear psyllid cadavers in Denmark 2016 and cultivated on solid media. In order to assess whether this (yet undescribed) species would be able to infect other insects, pathogenicity experiments were performed. Special regard was paid to psyllids (Hemiptera: Psyllidae). In the pathogenicity experiments, insects were inoculated with a “conidia shower”. For this purpose, mycelia mats were fixed to moist filter paper in the lid of small plastic cups, and a conidial shower was established. This is possible since fungi from Entomophthorales actively discharge their conidia [after Hajek et al. 2012]. A layer of agar on the bottom facilitated an increase in relative humidity above 95 % in the plastic cups and a filter paper on top prevented the psyllids from sticking to the agar. During 24 hours at 20 °C and short-day conditions (LD 10:14) the insects were exposed to the shower of the asexual conidia. The mortality as well as post mortal symptoms of the fungal infection were monitored over a period of 10 days after inoculation with Pandora sp. In our experiments, several Cacopsylla species vectoring distinct phytoplasmas to their respective host plants were successfully infected with this entomopathogenic fungus. The apple psyllid Cacopsylla picta Foerster, the vector of the cell wall lacking bacterium Candidatus Phytoplasma mali, the causing agent of apple proliferation disease, showed significantly increased mortality when inoculated with Pandora sp. compared to the control group. Some of the important vectors of phytoplasma diseases in European fruit production, C. pyri L., C. pyricola Foerster on pear and C. pruni Scopoli on Prunus sp. were also successfully infected and symptoms of the Pandora sp. infection on the cadavers were observed 24 hours after onset of mortality. The experiment showed that not only the adult stages of C. pyri were susceptible to the fungal infection, but also the nymphs of this pear psyllid. Other psyllids were also successfully infected with Pandora sp. such as C. peregrina Foerster feeding on hawthorn (Crataegus monogyna L.). Furthermore, other phloem-feeding hemipteran insects such as Acyrthosiphon pisum Harris (Hemiptera: Aphididae) vectoring several plant viruses, showed decreased survival probabilities after treatment with this entomopathogenic fungus. In the BMEL-supported research project “PICTA KILL”, Pandora sp. is being evaluated as a biocontrol agent for psyllid control. In an “Attract-and-Kill strategy” (s. Figure 1) this entomopathogenic fungus should be used as a “kill-component” in combination with specific attractant volatiles. In previous studies it has been shown that uninfected apple psyllids (C. picta) were more attracted to phytoplasma-infected apple trees, which emitted higher amounts of the sesquiterpene β-caryophyllene compared to uninfected plants (Mayer et al., 2008a). In bioassays, it was demonstrated that the synthetically produced β-caryophyllene was also highly attractive to the new generation of C. picta (Mayer et al. 2008b).

PheroFip 2019 83

Figure 1 - Attract-and-Kill strategy“ with encapsulated attractive volatiles and entomopathogenic fungus.

An innovative encapsulated formulation containing the entomopathogenic fungus Pandora sp. and volatiles such as β-caryophyllene should increase the specificity of this control approach and decrease the abundance of C. picta finally resulting in decreasing numbers of new infestations of apple trees with apple proliferation disease. We are currently designing experiments to elucidate this potential.

Key words: Psyllid control, entomopathogenic fungi, Apple proliferation, Cacopsylla picta, Pandora sp.

References Hajek, A. E., Papierok, B., Eilenberg, J. 2012: Chapter IX - Methods for study of the Entomophthorales. In: Manual of Techniques in Invertebrate Pathology (Second Edition) (eds. Lacey, L.A.): 285-316. Academic Press. Mayer, C. J., Vilcinskas, A., Gross, J. 2008 a: Phytopathogen Lures Its Insect Vector by Altering Host Plant Odor. J Chem Ecol. 34:1045–1049. Mayer, C. J., Vilcinskas, A., Gross, J. 2008 b: Pathogen-induced Release of Plant Allomone Manipulates Vector Insect Behavior. J Chem Ecol. 34: 1518–1522

PheroFip 2019 84

SESSION 6

APPLIED BIOTREMOLOGY: A NEW

DISCIPLINE FOR PEST MANAGEMENT.

VIBRATIONAL SIGNALS AS

SEMIOPHYSICALS

CHAIRPERSON: VALERIO MAZZONI

PheroFip 2019 85

Biotremology: from basic research to application

Virant-Doberlet M.1, Čokl A.1, Eriksson A.2, Lucchi A.3, Mazzoni V.2, Polajnar J.1

1Department of Organisms and Ecosystems Research, National Institute of Biology, Ljubljana, Slovenia; 2Research and Innovation Centre, Fondazione Edmund Mach, San Michele all'Adige (TN), Italy; 3Department of Agriculture, Food and Environment, University of Pisa, Pisa, Italy

Abstrat The increased awareness that substrate vibrations are an ancient and widespread form of animal communication, and that vibration receptors are ubiquitous in organisms, led to the establishment of biotremology as a new discipline of study of mechanical communication (Hill & Wessel, 2016). Surface-borne mechanical waves provide organisms with information about their environment that is crucial for their survival and reproduction. This information is not limited to intraspecific vibrational communication, but also includes prey detection, predator avoidance and information about habitat quality. While all insects possess highly sensitive vibration receptors, it is currently estimated that around 200,000 insect species use vibrational communication in a variety of intraspecific interactions (Cocroft & Rodríguez, 2005; Virant-Doberlet & Čokl, 2004). Vibrational signalling is particularly common in Hemiptera, which includes numerous major insect pests, like psyllids, leafhoppers, planthoppers and stink bugs. Reproductive behaviour in psyllids and stink bugs includes both, chemical and vibrational signals. However, in leafhoppers and planthoppers, mate recognition and location of the partner are mediated exclusively by substrate-borne vibrational signals (Čokl & Virant-Doberlet, 2003). During pair formation both sexes emit species- and sex-specific vibrational signals and vibrational interactions include continuous emission of vibrational songs and/or reciprocal exchange of male and female signals in a precisely coordinated duet. Competitive behaviour in males is expressed as emission of disruptive vibrational signals to interfere with ongoing duet and satellite behaviour, where intruding males silently approach the female duetting with another male (Mazzoni et al. 2009). As a result of a growing realisation of the ubiquitous nature of vibrations in the environment and about the importance of vibrational signals and cues in insect behavioural decisions, the interest to exploit substrate vibrations in pest management also increased in recent years (Čokl & Millar 2009; Mankin, 2012; Polajnar et al. 2015). Every movement of the insect body or its parts induces vibrations in the substrate and such incidental vibrations induced by walking and feeding can be used for monitoring. Detailed knowledge of the biology, ecology and behaviour of the target species is essential in order to exploit or manipulate insect behaviour. Current applications include the use of species-specific vibrational signals emitted in sexual communication for automatic detection (Korinšek et al., 2016) or for playback to attract insects to traps (Mazzoni et al. 2017) and interruption of mating behaviour by playback of natural or synthesized disruptive vibrational signals (Mazzoni et al., 2009). Although vibrational mating disruption is a novel approach (Eriksson et al. 2012), it has been already transferred to the field in the vineyards (Polajnar et al. 2016; Krugner & Gordon, 2018). Exploitation and manipulation of chemical signals is an integral part of IPM in several important crops and applied biotremology can provide innovative and efficient approaches in pest management of insects in which vibrational signals are an essential part of reproductive behaviour. Besides in depth studies of pest ecology and behaviour, implementation of such

PheroFip 2019 86

approach involves solving several technical challenges that include development of reliable and affordable sensors and playback devices, algorithms for automatic recognition of vibrational signals, as well as optimization of energy consumption, tailored to specific pests and specific field conditions. Taking into account that substrate vibrations are one of the most important and widespread sensory modalities guiding insect behavioural decisions, we believe that with more concentrated effort to solve current technical constraints, exploitation and manipulation of vibrational signals can be successfully included in IPM strategies and provide sustainable solutions in both, open field and greenhouse crop systems.

Key words: biotremology, substrate-borne vibrations, vibrational communication, Hemiptera, IPM

Acknowledgements The authors and activities were supported by the Slovenian Research Agency (research core funding no. P1-0255) (MVD, AČ, JP) and the EU 7th Framework Programme (grant agreement no. 265865), and by CBC Europe Ltd. (Milano, Italy) (AE, AL, VM).

References Cocroft, R. B. & Rodríguez, R. 2005: The behavioural ecology of insect vibrational communication. BioScience 55: 323-334. Čokl, A. & Millar J. C. 2009: Manipulation of insect signaling for monitoring and control of pest insects. In: Biorational Control of Arthropod pests (eds. Ishaaya, I. & Horowitz, A. R.): 279-316. Springer-Verlag, Heilderberg. Čokl, A. & Virant-Doberlet M. 2003: Communication with substrate-borne signals in small plant-dwelling insects. Annu Rev Entomol. 48: 29-50. Eriksson, A., Anfora, G., Lucchi, A., Lanzo, F., Virant-Doberlet, M., & Mazzoni, V. 2012: Exploitation of insect vibrational signals reveals a new method of pest management. PLoS ONE. 7: e32954. Hill, P. S. M. & Wessel, A. 2016: Biotremology. Curr Biol. 26: R187-R191. Korinšek, G., Derlink, M., Virant-Doberlet, M. & Tuma, T. 2016: An autonomous system for detecting and attracting leafhopper males using species- and sex-specific substrate-borne vibrational signals. Comput Electron Agric. 123: 29-39. Krugner, R. & Gordon, S. D. 2018: Mating disruption of Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae) by playback of vibrational signals in vineyard trellis. Pest Manag Sci. 74: 2013-2019. Mankin, R. W. 2012: Application of acoustics in insect pest management. CAB Rev. 7: 1-7. Mazzoni, V., Lucchi, A., Čokl, A., Prešern, J., & Virant‐Doberlet, M. 2009. Disruption of the reproductive behaviour of Scaphoideus titanus by playback of vibrational signals. Entomol Exp Appl. 133: 174-185. Mazzoni, V., Polajnar, J., Baldini, M., Rossi Stacconi, M. V., Anfora, G., Guidetti, R. & Maistrello, L. 2017: Use of substrate-borne vibrational signals to attract brown marmorated stink bug Halyomorpha halys. J Pest Sci. 90: 219-1229. Polajnar, J., Eriksson, A., Lucchi, A., Anfora, G., Virant-Doberlet, M. & Mazzoni, V. 2015: Manipulating behaviour with substrate-borne vibrations – potential for insect pest control. Pest Manag Sci. 71: 15-23. Polajnar, J., Eriksson, A., Lucchi, A., Virant-Doberlet, M. & Mazzoni, V. 2016: Mating disruption of a grapevine pest using mechanical vibrations: from laboratory to the field. J Pest Sci. 89: 909-921.

PheroFip 2019 87

Virant-Doberlet, M. & Čokl, A. 2004: Vibrational communication in insects. Neotrop Entomol. 33: 121-134.

PheroFip 2019 88

Vibrational Mating Disruption of Scaphoideus titanus: the current state after two years of field experience and benchmark analysis with Pheromonal Mating Disruption

Mazzoni V.1, Nieri R.2, Anfora G.1,3, Eriksson A.1, Lucchi A.4, Polajnar J.5, Virant-Doberlet M.5

1Research and Innovation Centre, Fondazione Edmund Mach, San Michele all'Adige (TN), Italy 2North Willamette Research and Extension Center, Oregon State University, Aurora (OR), United States; 3Centre Agriculture Food Environment, University of Trento, San Michele all'Adige (TN), Italy; 4Department of Agriculture, Food and Environment, University of Pisa, Pisa, Italy; 5Department of Organisms and Ecosystems Research, National Institute of Biology, Ljubljana, Slovenia

Abstract Biotremology is a new discipline that studies the vibrational communication in animals (Hill & Wessel, 2016). Vibrational mating disruption (VMD) is an innovative technique that is based on the transmission of vibrational signals into plant tissues to disrupt the mating behavior of insect pests (Eriksson et al. 2012). After the first laboratory experiments conducted in the second half of the 2000s (Mazzoni et al. 2009; Polajnar et al. 2016), the method has now been applied in commercial vineyards. In 2017, the world’s first vibrational vineyard was established in San Michele all’Adige (Northern Italy) for the control of the leafhopper Scaphoideus titanus. As a whole, we installed 110 emitters of disruptive noise on a surface of 1.5 ha of organic cabernet-franc. Each emitter was 50m distant from the next one. As a control, we used a nearby vineyard of the same variety and approximately the same size. Manual samplings were conducted throughout summers 2017 and 2018 at week periodicity from May to September. At each sampling, 20 randomly chosen leaves, from the median- basal part of 26 grapevines/plot were inspected to check the presence of leafhopper nymphs or other arthropods (namely, spiders or other no-target insects). The first results, at the second year of application, indicate a significant reduction from 2017 to 2018 of the number of S. titanus (<50%) and Empoasca vitis (<25%) nymphs in the vibrational vineyard as compared to the control. No effects on spiders and no-target insects were detected. To outline possible advantages and disadvantages of applying VMD to commercial crops, and for indicating weaknesses, strengths, similarities and differences, we made a benchmark analysis using the pheromone mating disruption (PMD) as reference. Both behavioral and physiological characteristics of the different target species and peculiarities of the two methods were included in this analysis: (1) sensorial modality and signal specificity; (2) searching behavior; (3) signal active space; (4) male rivalry behavior; (5) spatial dispersion; (6) insect phenology; (7) mechanisms of action; (8) efficacy assessment. We concluded that VMD is a promising method for control of pest species that rely on substrate borne vibrations for mating and that thanks to the constant and fast development of technologies (i.e. electronics, informatics, and energetics) it is quickly filling the gap to become applicable to commercial vineyards. In addition, we propose the use of the term “semiophysicals” as analogue of “semiochemicals” to indicate vibrational signals used for intra and interspecific communication in insects.

Key words: Biotremology, grapevine, leafhoppers, IPM

PheroFip 2019 89

Acknowledgements The activities were supported by the EU 7th Framework Programme (grant agreement no. 265865), and by CBC Europe Ltd. (Milano, Italy) that also built the prototype minishakers for field use in vineyards.

References Eriksson, A., Anfora, G., Lucchi, A., Lanzo, F., Virant-Doberlet, M. & Mazzoni, V. 2012: Exploitation of insect vibrational signals reveals a new method of pest management. PLoS One 7(3): e32954. Hill, P. S. & Wessel, A. 2016: Biotremology. Current Biology 26(5): R187-R191. Mazzoni, V., Lucchi, A., Čokl, A., Prešern, J. & Virant‐Doberlet, M. 2009: Disruption of the reproductive behaviour of Scaphoideus titanus by playback of vibrational signals. Entomologia experimentalis et applicata 133(2): 174-185. Polajnar, J., Eriksson, A., Virant-Doberlet, M. & Mazzoni, V. 2016: Mating disruption of a grapevine pest using mechanical vibrations: from laboratory to the field. Journal of pest science 89(4): 909-921.

PheroFip 2019 90

Exploiting vibrational communication for more efficient trapping of Halyomorpha halys (Heteroptera: Pentatomidae)

Jernej Polajnar1, Lara Maistrello2, Valerio Mazzoni3

1National Institute of Biology, Večna pot 111, Ljubljana, Slovenia; 2University of Modena and Reggio Emilia, Via Università 4, Reggio Emilia, Italy; 3Research and Innovation Centre, Fondazione Edmund Mach, Via E. Mach 1, San Michele all'Adige (TN), Italy

Abstract Halyomorpha halys, or the Brown Marmorated Stink Bug (BMSB) is commonly recognized as one of the most notorious invasive insect species. After the population outbreak in some areas of North America, a locally severe population outbreak is also occurring in the north Italy’s Po valley (Maistrello et al. 2018), where BMSB has quickly become a key pest in fruit orchards (Maistrello et al. 2017). Its invasion potential is a cause of great concern among farmers and plant protection services in areas with suitable climate worldwide (Kriticos et al. 2017). Control measures against H. halys currently consist of frequent applications of broad- spectrum insecticides, which is adding to environmental pesticide burden and disrupting IPM programs in affected areas (Leskey et al. 2012). Control efforts are hampered by incredible polyphagy of H. halys, its high dispersion potential, high reproductive potential and general robustness. Aside from biological control, behavioral manipulation using attraction to male- emitted sexual pheromones is one of the more promising approaches. The male-emitted aggregation pheromone has been identified recently (Khrimian et al. 2014) and is commercially available, although pheromone attraction of H. halys has the same shortcoming as in other species of stink bugs: traps’ efficiency is more suited for monitoring than mass trapping purpose (Morrison et al. 2015) because attraction to the pheromone source is not precise in this family. The likely reason for low efficiency of pheromone traps is that communication in stink bugs is bimodal. Pheromones attract males, females, and late-instar nymphs to general vicinity of the emitter (Khrimian et al. 2014), but the final approach during the courtship of stink bugs is mediated by substrate-borne vibrational signals (Virant-Doberlet & Čokl, 2004). Thus, we argue that mechanical vibrations can be used as a “bridge” connecting long-range attraction towards the source of synthetic pheromones and actual elimination. To explore this possibility, an ongoing research effort was started in 2015, beginning with the basic description of vibration-mediated sexual behaviour of H. halys. Courtship involved a stereotypical exchange of signals between a male and a female which started with the male spontaneously producing vibrational pulses (labeled MS-1), to which receptive females replied. A duet was formed, characterized mainly by sequences of shorter, regularly repeated female signals (FS-2), to which males reacted by searching for the source. Both partners emitted other types of signals during this time as well, ending with copulation (Polajnar et al. 2016). We then performed a series of laboratory trials in which we reproduced male and female signals using an electromagnetic “shaker”. In particular, we tested the potential aggregation function of the long, spontaneously produced MS-1 signals, and the male attraction to the regularly repeated FS-2 signals. We did not observe any effect of MS-1 playback, but FS-2 signals were attractive to adult males on several types and geometries of substrate (both natural and artificial). We observed a so-called “loitering effect”, where the male circled

PheroFip 2019 91

continuously around the exact location where the minishaker was attached to the experimental arena, and never left the area (Mazzoni et al. 2017). Thus, the FS-2 was selected for field trials. A trap prototype was constructed (Biogard®, CBC Europe S.r.l.), implementing a commercial aggregation pheromone dispenser, an electromagnetic “shaker” continuously vibrating the whole trap with the FS-2 sequence, and a circuit emitting electric shocks on top of the standard pyramid panel construction. The males were attracted to the trap by the pheromone and then driven inside cavities in its head, where they were killed by electrocution. Preliminary results show highly male-biased capture rates, thus proving the effect of vibrational playback per se. Further work in the following seasons will focus on improving capture rate which is crucial if the devices are to be used in mass trapping of H. halys. In particular, attractiveness to adult males only needs to be compensated to achieve meaningful reduction of population density.

Key words: biotremology, invasive pest, behavioural manipulation, multimodal trap, IPM

Acknowledgements The work was made possible by the grant from ‘Fondazione Cassa di Risparmio di Modena’ (2013.065) and by CBC Europe Ltd. (Milano, Italy). Presenting author’s participation is also supported by the Slovenian Research Agency (research core funding no. P1-0255).

References Khrimian, A., Zhang, A., Weber, D. S., Ho, H.-Y., Aldrich, J. R. et al. 2014: Discovery of the aggregation pheromone of the brown marmorated stink bug (Halyomorpha halys) through the creation of stereoisomeric libraries of 1-bisabolen-3-ols. J. Nat. Prod. 77: 1708-1717. Kriticos, D. J., Kean, J. M., Phillips, C. B., Senay, S. D., Acosta, H., Haye, T. 2017: The potential global distribution of the brown marmorated stink bug, Halyomorpha halys, a critical threat to plant biosecurity. J. Pest Sci. 90: 1033-1043. Leskey, T. C., Short, B. D., Butler, B. R., Wright, S. E. 2012: Impact of the invasive brown marmorated stink bug, Halyomorpha halys (Stål), in mid-Atlantic tree fruit orchards in the United States: Case studies of commercial management. Psyche 2012: 535062 Maistrello, L., Vaccari, et al. 2017: Monitoring of the invasive Halyomorpha halys, a new key pest of fruit orchards in northern Italy. J. Pest Sci. 90: 1231-1244. Maistrello, L., Dioli, P., Dutto, M., Volani, S., Pasquali, S., Gilioli, G. 2018: Tracking the spread of sneaking aliens by integrating crowdsourcing and spatial modelling: the Italian invasion of Halyomorpha halys. BioScience, in press. Mazzoni, V., Polajnar, J., Baldini, M., Rossi Stacconi, M. V. Anfora, G. et al. 2017: Use of substrate-borne vibrational signals to attract the brown marmorated stink bug, Halyomorpha halys. J. Pest Sci. 90: 1219-1229. Morrison, W. R. III., Cullum, J. P., Leskey, T. C. 2015: Evaluation of trap designs and deployment strategies for capturing Halyomorpha halys (Hemiptera: Pentatomidae). J. Econ. Entomol. 108: 1683-1692. Polajnar, J., Maistrello, L., Bertarella, A., Mazzoni, V. 2016: Vibrational communication of the brown marmorated stink bug (Halyomorpha halys). Physiol. Entomol. 41: 249-259. Virant-Doberlet, M.; Čokl, A. 2004: Vibrational communication in insects. Neotrop. Entomol. 33: 121-134.

PheroFip 2019 92

SESSION 7

INVASIVE PESTS: A CHALLENGE FOR IPM

A - BROWN MARMORATED STINK BUG AND

OTHER INVASIVE HEMIPTERANS

CHAIRPERSON: GREG KRAWCZYK

PheroFip 2019 93

The invasive Halyomorpha halys in Europe: a challenge for integrated fruit production

Lara Maistrello1

1Department of Life Sciences, University of Modena and Reggio Emilia, Via G. Amendola 2, 42122, Reggio Emilia, Italy

Abstract The Brown Marmorated Stink Bug (BMSB), Halyomorpha halys Stal (Heteroptera, Pentatomidae), is a highly polyphagous insect native to eastern Asia, rapidly spreading as a serious pest of global importance for many agricultural crops (Haye & Weber, 2017). It is also a household nuisance, due to the large overwintering aggregations inside man-made structures. It has successfully invaded and established in large areas of North America, in many central, eastern and southern European countries and, most recently, also in Chile (Leskey & Nielsen, 2018). BMSB caused millions of dollars losses in fruit orchards and horticultural crops in the USA, it rapidly became a key pest of fruit orchards in Northern Italy with severe losses especially on pear (Maistrello et al. 2017) and is a serious pest of hazelnuts grows in Piedmont and in the countries along the Black Sea (Bosco et al. 2017). The sneaking behaviour of BMSB makes its worldwide invasion virtually unstoppable. In fact, the propensity to aggregate in narrow, hidden microhabitats (e.g. slots of vehicles, packaging materials, suitcases, clothes etc.) exhibited particularly during overwintering, allows these bugs to “hitchhike” unnoticed on practically any type of goods, highly facilitating human assisted spread all over the world (Maistrello et al. 2018). Interceptions of BMSB are increasingly reported at transitional facilities in many countries, and models highlight the potential for further spread and establishment, threatening horticultural productions of areas with suitable climate in both Hemispheres (Kriticos et al. 2017). BMSB is currently rapidly spreading in European countries causing increasing damage in fruit orchards and other horticultural crops. In northern Italy a life table study performed outdoors demonstrated the high growth potential of BMSB bivoltine populations with high reproductive rates for both generations (R0 = 24.04 and 5.44 respectively) (Costi et al. 2017). A 3-year field survey in the same area showed that the biocontrol potential by native antagonists is presently very limited (Costi et al. 2018). In the attempt to counteract the fast spread and high damage levels caused by BMSB, farmers intensify treatments with broad-spectrum insecticides, resulting in disruption of the existing Integrated Pest Management (IPM) programs (Leskey et al. 2012; Maistrello et al. 2017), with increasing negative environmental impact. Management of BMSB is particularly challenging. Chemical control proved to be scarcely effective due to the general robustness, as well as to the high polyphagy combined with high mobility of all instars, that results in continuous movements of the bugs between different host plants (Lee & Leskey, 2015). More sustainable approaches to manage BMSB in fruit orchards include the use of exclusion netting systems (Caruso et al. 2017) are behaviour-based strategies such as IPM-CPR (Blaauw et al., 2015). Evaluation of mass rearing of efficient native antagonists for inundative release is also being considered. Field monitoring of BMSB presently relies on traps baited with aggregation pheromones that showed scarce efficiency and increased damage on the plants surrounding the installation point. The use of substrate-borne vibrational stimuli might favour the development of

PheroFip 2019 94

innovative multi-modal trapping devices as well as other behavioural manipulation approaches (Mazzoni et al. 2017).

Key words: brown marmorated stink bug, biological invasions, human assisted spread, fruit orchards, IPM

Acknowledgements The work was partially funded by a grant from ‘Fondazione Cassa di Risparmio di Modena’ (2013.065) and by the Emilia Romagna region, within the Rural Development Plan 2014- 2020 Op. 16.1.01- GO PEI-Agri - FA 4B, Pr. «HALYS», coordinated by CRPV.

References Blaauw, B. R, Polk, D. & Nielsen, A. L. 2014: IPM-CPR for Peaches: Incorporating Behaviorally-Based Methods to Manage Halyomorpha halys and Key Pests in Peach. Pest Manag. Sci. 71: 1513–22. Bosco, L., Moraglio, S. T. & Tavella, L. 2017: Halyomorpha halys, a serious threat for hazelnut in newly invaded areas. J Pest Sci. 91: 661-670. Caruso, S., Vaccari G., Vergnani S., Raguzzoni, F. & Maistrello, L., 2017. Nuove opportunità di impiego di reti multifunzionali. L’Informatore Agrario 15: 57-60. Costi, E., Haye, T. & Maistrello, L. 2017: Biological parameters of the invasive brown marmorated stink bug, Halyomorpha halys, in southern Europe. J Pest Sci 90: 1059– 1067. Haye, T. & Weber, D. C. 2017: Special issue on the brown marmorated stink bug, Halyomorpha halys: An emerging pest of global concern. J. Pest Sci. 90: 987-988. Costi, E., Haye, T. & Maistrello, L. 2018: Egg parasitization and predation of the invasive Halyomorpha halys by native antagonists: a three-year field survey in Northern Italy. J. Appl. Entomol. In press. Kriticos, D. J., Kean, J. M., et al. 2017: The potential global distribution of the brown marmorated stink bug, Halyomorpha halys, a critical threat to plant biosecurity. J. Pest Sci. 90: 1033-1043. Lee, D. H. & Leskey, T. C. 2015: Flight behavior of foraging and overwintering brown marmorated stinkbug, Halyomorpha halys (Hemiptera: Pentatomidae). Bull. Ent. Res.105: 566-73. Leskey, T. C. & Nielsen, A. L. 2018: Impact of the Invasive Brown Marmorated Stink Bug in North America and Europe: History, Biology, Ecology, and Management. Annu. Rev. Entomol. 63: 599-618. Mazzoni, V., Polajnar, J. et al. 2017: Use of substrate-borne vibrational signals to attract the brown marmorated stink bug, Halyomorpha halys. J. Pest Sci. 90: 1219-1229. Maistrello, L., Vaccari, G. et al. 2017: Monitoring of the invasive Halyomorpha halys, a new key pest of fruit orchards in Northern Italy. J. Pest Sci 90: 1231-1244. Maistrello, L., Dioli et al. 2018: Tracking the spread of sneaking aliens by integrating crowdsourcing and spatial modelling: the Italian invasion of Halyomorpha halys. BioScience, in press.

PheroFip 2019 95

Agroecosystem Impacts on Brown Marmorated Stink Bug Pheromone Trap Capture and Damage in North Carolina Apple Orchards

James Walgenbach1, Steven Schoof1, Javier Gutierrez2, David Crowder2

1Mountain Horticultural Crops Research & Extension Center, NC State University, Mills River, NC 28759; 2Department of Entomology, Washington State University, Pullman WA 99164.

Abstract The brown marmorated stink bug (BMSB), Halyomorpha halys (Stål), is an invasive pest of Asian origin that was first detected in the US in the state of Pennsylvania in the mid 1990s. By 2010 it had spread approximately 700 km south to North Carolina, and by 2015 populations were causing economic damae in the central piedmont and western mountainous regions of the state. It is currently a major pest of apples, peaches, fruiting vegetables, as well as corn and soybeans in these regions of the state. The need for pyrethroid insecticides to control this pest has severely disrupted long-standing apple IPM programs that relied on mating disruption, narrow-spectrum insecticides, and biological control as management tools, and has led to outbreaks of European red mite, woolly apple aphid, and San Jose scale. Studies were conducted in commercial apple orchards in 2016 and 2017 using pheromone traps (BMSB pheromone + MDT synergist) and damage assessments to determine the phenology and distribution of BMSB populations and damage in apple orchards. Key findings from these studies included; 1) There were no differences in capture of adults in pheromone traps placed in the exterior versus interior of orchards, but nymphal captures were significantly higher in exterior traps; 2) When examined by month (June, July, August and September), the only instance where significant relationships existed between pheromone trap captures and damage was in June; and 3) When damage was assessed along transects from the edge to 50 meters into orchards, damage was significantly higher on the edge row, which was also within 5 m of pheromone traps placed on the exterior of orchards. These results raise several questions about apple agroecosystem effects on BMSB pheromone trap captures and damage within apple orchards in the Southern Appalachian Mountains of western NC. The lack of significant relationships between pheromone trap captures and damage to apples complicates the interpretation of trap captures and decisions on where to spatially deploy traps in orchards. Also, the lack of differences in captures of bugs in traps placed in the exterior and interior of orchards may suggest agroecosystem factors are affecting the spatial distribution of BMSB trap captures and damage within orchards. In 2018, studies were conducted to characterize the impact of several agroecosystem effects on BMSB pheromone trap capture and damage in apple orchards, including adjacent habitat, orchard size, cultivar, and pest management practices. The relationship between pheromone trap captures and damage was also assessed. These studies included 25 commercial apple orchards that varied in size and surrounding habitat (e.g., wooded habitats, row crops, pastures, buildings), factors that may affect the spatial and temporal distribution of BMSB in apples. Two parallel sampling transects separated by 50 m were established in each orchard to monitor damage along a gradient from the edge to the center of the orchard; one transect contained pheromone traps at the sampling sites and one did not have traps. The habitat adjacent to the border sample site was the same for both transects, but varied among orchards. Additional data collected included cultivar, tree height, BMSB insecticide sprays, and other orchard borders. A landscape level multicomponent analysis was conducted to assess the impact of these factors on BMSB trap capture and damage.

PheroFip 2019 96

While analysis of 2018 data is not yet complete, results showed that the strongest correlation to both BMSB pheromone trap captures and damage to apples was time, with both increasing in successive months from June through September. These results are consistent with the phenology of BMSB in NC, where overwintering adults emerge from late April through May, and first generation adults emerge from late July through August, with peak adult trap captures in mid September. Previous studies have shown that non-managed wooded habitats are key reservoirs of reproducing populations of BMSB, and it was hypothesized that they served as key sources of adults infesting apple orchards, and also that BMSB damage to fruit would decline along a gradient from the orchard edge adjacent to a wooded habitat to the center of the orchard. However, there was no significant relationship between pheromone trap capture and adjacent habitat, or between pheromone trap capture and location of trap from the edge of the orchard. The most striking observation was that fruit damage along transects into orchards was dramatically higher in transects containing pheromone traps (40.1% damage) versus no traps (1.0%). Also, there was only a weak correlation between trap capture and distance into the orchard. These results suggest that BMSB dispersal into apple orchards in the highly diverse habitats of the southern Appalachian Mountains may occur on a scale that is sufficiently large to negate the detection of habitat or edge effects in apple orchards. Apple orchards are relatively small in this region (average size of about 8 ha), and may be less susceptible to nearby habitat effects or dispersal gradients compared to regions with larger orchards. Also, the strong attraction of pheromone lures to BMSB was evident in that high levels of fruit damage were only observed where pheromone traps were located. This bodes well for the use of pheromone lures in attract and kill approaches to managing BMSB.

Key Words: Brown marmorated stink bug, apples, pheromone traps, sampling

Acknowledgments The authors thank cooperating growers for use of their orchards and sharing of insecticide spray records. This research was supported, in part, by USDA-NIFA SCRI project 2016- 51181-25409.

PheroFip 2019 97

Biocontrol of the invasive Halyomorpha halys

Hillary Peterson, Jared Ali, Greg Krawczyk

Abstract The brown marmorated stink bug (BMSB), Halyomorpha halys (Stål), is an invasive polyphagous insect pest originating from Asia. It has disrupted successful integrated pest management programs in fruit due to the need for an intensive use of broad-spectrum insecticides for effective control, often leading to secondary outbreaks of other orchard pests such as mites, aphids, and scales. A necessary step to reestablish these once effective IPM programs is to understand which natural enemies are influencing populations of BMSB. Since the major outbreak of BMSB in the Mid-Atlantic region of the United States in 2010, several research groups have conducted sentinel surveys to determine predation and parasitism of BMSB across agricultural landscapes. These surveys have found native hymenopteran egg parasitoids, including members of the families Scelionidae and Eupelmidae, attacking BMSB in low numbers, indicating the ability of these native parasitoids to find the eggs of the invasive species. Additionally, BMSB sentinel egg surveys in Maryland in 2014 discovered an adventive population of Trissolcus japonicas (Ashmead), one of the most successful egg parasitoids of BMSB in its native range of Asia. The aims of this study were to understand which species of native egg parasitoids attack BMSB, to determine if T. japonicus populations are present and growing in commercial orchards and surrounding landscapes in Pennsylvania, and to understand which landscape factors play a role in the parasitism of BMSB eggs. Sentinel egg masses were deployed from June through August in 2016, 2017, and 2018. Egg masses were collected fresh (<48 hours since laid) from colonies at the Penn State Fruit Research and Extension center (Biglerville, PA), or purchased from an indoor colony (New Jersey Department of Agriculture). Egg masses were deployed on the underside of leaves of orchard trees and known BMSB host plants in wooded landscapes surrounding the orchard, and on the border between the orchard and woods. Eggs were deployed as fresh or frozen, and either left on the leaves of colony host plants, or affixed as to a white square of card stock paper using double sided sticky tape. Egg masses were deployed for up to 72 hours before collection, and allowed at least six weeks in a growth chamber for parasitoids to emerge. All emerged parasitoids were identified to species, and rates of parasitism were calculated. Egg masses were dissected after six weeks to determine rates of partially developed parasitoids. In addition to these deployments, egg masses found in the landscape laid by native stink bug species and BMSB were collected to determine natural rates or parasitism. In 2017 and 2018, additional surveys for parasitoids were conducted utilizing yellow sticky cards baited with a BMSB aggregation pheromone. Finally, in 2018, field sites were monitored for the presence of flowering plants, in order to determine if these food sources for the parasitoids may be associated with higher rates of parasitism. In all three years, species within the families Scelionidae and Eupelmidae were found to attack BMSB egg masses in the orchard and surrounding landscapes. In 2017, T. japonicas populations were detected in two Pennsylvania counties on yellow sticky cards. In 2018, T. japonicus populations were detected in two additional Pennsylvania counties on yellow sticky cards, and also were reared from both sentinel egg masses and naturally laid BMSB egg masses. In all three years, less than 5% of sentinel egg masses were parasitized, however, collected egg masses (including native stink bug eggs) had a much higher rate, with over 55% of naturally laid egg masses being parasitized. Understanding when and where these egg

PheroFip 2019 98

parasitoids are attacking BMSB, and how they find the eggs, will aid in determining the how to conserve these biological control agents.

Key words: brown marmorated stink bug, Halyomorpha halys, egg parasitoid, Trissolcus japonicus

PheroFip 2019 99

Predatory ability of wild generalist predators on eggs and first instars of Halyomorpha halys

Giacomo Bulgarini1, Zaid Badra2, Lara Maistrello1

1Dipartimento Scienze della Vita, Università di Modena e Reggio Emilia, Via G. Amendola 2, 41122, Reggio Emilia, Italy; 2Department of Mountain Environment and Agriculture, Free University of Bolzano, Piazza Università 5, 39100, Bolzano, Italy

Abstract Halyomorpha halys (Heteroptera, Pentatomidae), native to East Asia, is a polyphagous species with more than 300 host plants recognized (Rice et al. 2014). Thanks to its close association with manmade structures during the overwinter period, that facilitates human assisted spread (Maistrello et al. 2018), this insect is a fast spreading invasive pest of fruit orchards and many other crops both in the U.S.A. (Leskey & Nielsen, 2018) and in Italy (Maistrello et al. 2017). Among the direct damage that this insect can cause there are malformation, suberisation, discoloration, necrotic areas and watery rot, all damage that render products unmarketable (Rice et al. 2014). To manage the invasion of this pest, farmers have increased the use of broad spectrum insecticides, resulting in disruption of the most innovative IPM strategies, with serious risks both for the economy and the environment (Maistrello et al. 2017, Leskey et al. 2012). In fact the use of broad spectrum insecticides can also lead to the reduction of beneficial insects, such as pollinators, parasitoids and predators, altering the natural balance. Therefore, it is necessary to identify more sustainable strategies to control this pest. H. halys is an alien species for Italy, thus species specific antagonists are lacking in the newly invaded areas. However, native species of generalist predators and parasitoids could eventually use H. halys as prey/host, becoming good candidates for the biological control. As matter of fact, generalist predators can be more effective on invasive species than the specialist predators in their introduced range (Chang & Kareiva, 1999). In the present study, the predatory ability of wild collected European native generalist insect predators was verified on eggs and young instars of H. halys using a no choice test. For the study, the following European native species were tested: Forficula auricularia (Dermaptera, Forficulidae), Harmonia axyridis (Coleoptera, Coccinellidae), Pholidoptera littoralis (Orthoptera, Tettigonidae), Nagusta goedelii (Rhyncota, Reduviidae), Rhynocoris iracundus (Rhyncota, Reduviidae) and two species of the Himacerus genus (Rhyncota, Nabidae). These predators were captured in green areas and urban parks of Reggio Emilia (Northen Italy) using tree beating and sweep net techniques. After a starving period of 24 hours, each predator was individually placed inside a transparent box with a bean plant bearing at least two well developed leaves and the H. halys prey item, that consisted of one egg mass or a hatched egg mass (with first instar nymphs) or 5 second instar individuals. R. iracundus was also tested with adults of H. halys. For each replicate, a control was carried out, with the same set up but without the presence of the predator. After 48 hours the survival of the prey items and the predators was recorded, and, for the egg masses, emergence was checked during the following 5 days. From the comparison between the survivors of the treatment group (prey and predator) and the control one (no predators) it emerged that Pholidoptera littoralis is the only predator capable of predating all the instars tested, including eggs. The other predators did not feed on the eggs, although F. auricularia damaged them. N. goedelii significantly predated the first

PheroFip 2019 100

instars, while R. iracundus and nabids of the Himacerus genus preyed on second instars. R. iracundus significantly predated also adult individuals of H. halys. This study showed the potential of some European native generalist predators for the biological control of H. halys, in particular of R. iracundus and P. littoralis. The augmentative release of these predators into cultivated areas could negatively affect the survival of H. halys in the fields, and especially in combination with augmentative release of native parasitoids, such as Anastatus bifasciatus (Hymenoptera, Eupelmidae) (Costi et al. 2018), it might result in increased impact and effectiveness of biological control against this invasive pest. However further research is necessary both in the laboratory (preference tests) and in the field to verify their effectiveness and the possible consequences on the agroecosystem.

Acknowledgements The work was partially funded by the Emilia Romagna region, within the Rural Development Plan 2014-2020 Op. 16.1.01- GO PEI-Agri - FA 4B, Pr. «HALYS», coordinated by CRPV.

References Chang, G. C. & Kareiva, P. 1999: The case for indigenous generalists in biological control. In: Theoretical Approaches to Biological Control 103–115. Costi, E., Haye, T. & Maistrello, L. 2018: Egg parasitization and predation of the invasive Halyomorpha halys by native antagonists: a three year field survey in Northern Italy. Journal of Applied Entomology. In press. Leskey. T. C., Hamilton, G. C., Nielsen, A. L., Polk, D. F., Rodriguez, Saona C., Bergh, J. C., Herbert, D. A., Kuhar, T. P., Pfeiffer, D., Dively, G. P., Hooks, C. R. R., Raupp, M. J., Shrewsbury, P. M., Krawczyk, G., Shearer, P. W., Whalen, J., Koplinka Loehr C., Myers, E., Inkley, D., Hoelmer, K. A., Lee, D. H. & Wright, S. E. 2012: Pest Status of the Brown Marmorated Stink Bug, Halyomorpha halys in the USA. Outlooks Pest Manag. 23: 218–226. Leskey, T. C. & Nielsen, A. L. 2018: Impact of the Invasive Brown Marmorated Stink Bug in North America and Europe: History, Biology, Ecology, and Management. Annu. Rev. Entomol. 63: 599 618. doi: 10.1146/annurev ento 020117 043226. Maistrello, L., Vaccari, G., Caruso, S., Costi, E., Bortolini, S., Macavei, L., Foca, G., Ulrici, A., Bortolotti, P. P., Nannini, R., Casoli, L., Fornaciari, M., Mazzoli, G. L. & Dioli, P., 2017: Monitoring of the invasive Halyomorpha halys, a new key pest of fruit orchards in Northern Italy. Journal of Pest Science 88(1): 37–47. DOI 10.1007/s10340 017 0896 2. Maistrello, L., Dioli, P., Dutto, M., Volani, S., Pasquali, S. & Gilioli, G. 2018: Tracking the spread of sneaking aliens by integrating crowdsourcing and spatial modelling: the Italian invasion of Halyomorpha halys. BioScience, in press. Rice, K. B., Bergh, C. J., Bergmann, E. J., Biddinger, D. J., Dieckhoff, C., Dively, G. & Herbert, A. 2014: Biology, ecology, and management of brown marmorated stink bug (Hemiptera: Pentatomidae). Journal of Integrated Pest Management 5(3): A1 A13

PheroFip 2019 101

Feeding, Mating, and Flight Dispersal Behavior of the Invasive Pest Species Lycorma delicatula (Hemiptera: Fulgoridae)

Thomas C. Baker1*, Andrew J. Myrick1

1Department of Entomology, Center for Chemical Ecology, Penn State University, University Park, Pennsylvania, USA *[email protected]

Abstract The spotted lanternfly, Lycorma delicatula, is an invasive fulgorid bug species that was recently accidentally introduced into North America, likely from indigenous populations in China. The first record of this species in the United States occurred in September, 2014 in Berks County, Pennsylvania. This incipient pest of vineyards, fruit orchards, and hardwood forests of the U.S. has now spread from a quarantine zone in a small portion of one county in eastern Pennsylvania in 2015 to now comprise in 2018 over 6,900 square miles of infestation, with recent detections reported also in two neighboring states – Delaware and New York. There is great concern of this species’ certain continued spread over a broader geographic area in North America if it cannot be controlled. In 2017 we saw a progression of adult behaviors that culminated in a mass flight dispersal of thousands of adults followed by mating and egg laying. After one to two weeks of intense early-season feeding, short flights occurred indiscriminately from the foliage of one tree to another or between bushes, trees and vines. At the peak of adult population abundance in 2017 these flights became more prevalent and lengthier, with adults of both sexes observed to launch themselves into the wind from all types of host and non-host trees or from porches, posts and other human-made structures. The fairly straight-line level or gradually descending flight trajectories allowed the flying adults to traverse only usually 30 to 80 meters of ground in one episode, showing no preference for landing on any particular species of plant or inanimate object. They would then crawl up a nearby vertical object and launch themselves into flight again. Thousands of adult L. delicatula undertook such flights together on any day, creating massive swarms in local areas. We developed a novel technique to study the in-flight wind-orientation of adults during their flight dispersal bouts.

PheroFip 2019 102

Alternative methods to manage brown marmorated stink bug, Halyomorpha halys

Greg Krawczyk1*, Hillary Morin1, Claire Hirst1

1The Pennsylvania State University, Department of Entomology, Fruit Research and Extension Center, Biglerville, PA 17307 *[email protected]

Abstract As the insecticide treatments remain the most effective tools to manage the brown marmorated stink bug (BMSB) Halyomorpha halys (Stäl) (Heteroptera - Pentatomidae) in Pennsylvania fruit orchards, we also attempt to develop alternative methods to manage this pest in more sustainable, environmentally responsible way. The BMSB monitoring studies from previous seasons, which included lure and trap design comparisons, documented the viability of BMSB field monitoring practices for accurate assessment of the actual BMSB pest pressure at least from mid-July until October. During the 2017 and 2018 seasons we tested deltamethrin insecticide treated nets (D-Terrence net, Vestergaard Group, Lausanne, Switzerland) baited with BMSB monitoring lures (3x or 5x) either from Trece Inc. (Adair, OK) or AgBio Inc. (Westminster, CO) to monitor BMSB populations around commercial apple orchards. Starting from late July, the treated nets were placed on 2.4 m high shepherd hooks and distributed outside of the apple blocks, at about 50 m distance between each net, however higher number of nets were placed in the direction of potential influx of BMSB from outside vegetation such as woods. To assess the number of BMSB adults and nymphs killed by nets, plastic tarps 1.8 m in diameter were positioned under some net stations. The standard BMSB monitoring sticky traps (Trece Inc.) or container based Rescue traps (Sterling Int.) were used to assess the pest population in the orchards surrounded by the net traps and control blocks. At the peak BMSB activity periods high numbers of BMSB adults and nymphs, as well as native stink bugs were collected from tarps under the BMSB net traps. The numbers of BMSB captured by standard monitoring traps located inside the orchards surrounded by net traps were always lower than in the control apple blocks. Although the practical implications of our experiment with insecticide nets still remains to be determined, the practical results from our trials are very promising and potentially can be used for the development of alternative BMSB trapping method(s).

Key words: Halyomorpha halys, insecticide treated nets, BMSB monitoring, pome fruit, stone fruit

PheroFip 2019 103

SESSION 7

INVASIVE PESTS: A CHALLENGE FOR IPM

B - SPOTTED WING DROSOPHILA AND OTHER INVASIVE

DIPTERANS

CHAIRPERSON: HOWARD THISTLEWOOD

CHAIRPERSON: HEIDRUN VOGT

PheroFip 2019 104

Automatic identification of Drosophila suzukii (Spotted Wing Drosophila, SWD) by a sensor measuring the wingbeat frequency

Tim Belien1, Yenisey M. Mendoza2, Bart De Ketelaere2, Ammar Alhmedi1, Frank Mathijs2, Wouter Sayes2, Rik Clymans1, Dany Bylemans1

1pcfruit, Zoology Department, Sint-Truiden, Belgium 2Katholieke Univ Leuven, Div Mech Biostat & Sensors MeBioS, Leuven, Belgium

Abstract The Asiatic vinegar fly Drosophila suzukii (Spotted Wing Drosophila, SWD) has recently invaded the European and American continent, and immediately became a key pest of a wide variety of soft-skinned fruit crops. Effective monitoring and early detection of D. suzukii is crucial for appropriate management actions against this devastating pest. The monitoring of D. suzukii is currently primarily executed by manually inspection of installed traps with liquid apple cider vinegar and yeast-based lures. However, many growers are unable to distinguish the harmful D. suzukii flies from other harmless Drosophila species in monitoring traps. This is detrimental for IPM as broad spectrum insecticides are often sprayed preventively at the first Drosophila sp. notice. Automatic monitoring traps for D. suzukii would overcome this problem by measuring, identifying and counting D. suzukii flies as they enter the trap, and providing clear information concerning the presence of D. suzukii to the grower. Therefore, we developed a sensor-based system capable of identifying D. suzukii based on the measurement of its wingbeat characteristics. In this approach an optoelectronic sensor is used that records the wingbeat of an entering flying insect. In laboratory experiments we demonstrated that using this sensor D. suzukii can be identified and distinguished from the common fruit fly D. melanogaster. Furthermore we investigated the influence of the age, gender, temperature and relative humidity on the measured wingbeat frequencies for both fruit fly species. Temperature and relative humidity did have an impact on the measured wingbeat frequency signal, indicating the need for measuring these environmental data as well in the automatic monitoring trap. Also the age of the flies did have an influence on the wingbeat measurements. The practical consequences of these findings for the further field validation of this optoelectronic wingbeat sensor are discussed.

Key words: Drosophila suzukii, optoelectronic wingbeat sensor, automatic monitoring

PheroFip 2019 105

Comparative study of Drosophila suzukii females’ behavioral responses to fruit odors of two varieties of Vitis vinifera

Amani Alawamleh1*, Gordana Ðurović2, Maria Giovanna Di Stefano1, Sonia Ganassi1, Massimo Mancini1, Felix Wäckers3, Gianfranco Anfora2, Antonio De Cristofaro1

1Department of Agricultural, Environmental and Food Sciences, University of Molise, Via De Sanctis, Campobasso, Italy; 2Fondazione Edmund Mach, Via Edmund Mach 1, San Michele all’Adige, Trento, Italy.3Biobest Group NV, Ilse Velden, Westerlo, Belgium. * [email protected]

Abstract We report olfactometer results on comparative study of Drosophila suzukii females’ behavioral responses to fruit odors of two varieties of Vitis vinifera var.Vittoria and Black magic.

Introduction Drosophila suzukii Matsumura (Diptera: Drosophilidae), is a highly polyphagous pest of fresh fruits. It is the only Drosophilid fly with serrated ovipositor, which enables it to oviposit in unwounded fresh fruits, thereby making them unmarketable (Cini et al. 2012). Recently, D. suzukii invaded Europe and is heavily affecting fruit industry (Rota-Stabelli et al. 2013). Numerous evidences have shown the importance of host plant volatile compounds in oviposition site-selecting behavior by insect herbivores. Recent studies showed that the microbes (yeasts and bacteria) play a vital role in the above process (Becher et al. 2012). At present, not so efficient monitoring and control tools of D. suzukii have been developed. The present study aims to evaluate the behavioral responses of D. suzukii females to odors released by fresh and ripened grapes fruits i.e. Vitis vinifera, using behavioral bioassays (Y-tube olfactometer), which is expected to contribute in the designing of innovative concepts for plant protection technologies and development of trapping systems for the integrated pest management strategies against this pest.

Material and methods Drosophila suzukii colony was reared on a standard Drosophila semi-artificial diet at 24±1 °C, 65±5% relative humidity (RH), and a photoperiod of 16:8 (L:D) h. Seven days old adult females were starved for 1 h in 200 ml plastic container on water soaked cotton swab to enhance the fly response towards odors. Only naive mated females were used in the study and the fruits tested were two different varieties of table grapes, Vitis vinifera var. Vittoria (white grapes) and Black magic (black grapes), that were freshly picked from different fields in Apulia region, Italy, during July-August 2018, and kept at room temperature (25°C) for 1 h before conducting the experiments. The response of D. suzukii females to host fruit odors was investigated in two-choice assays using Y-tube olfactometer (stem 29 cm, arm length 22 cm, arm angle 60°, internal diameter 4.5 cm) one arm of Y-tube hold 30 g of fresh grapes fruits which were randomly assigned to avoid positional biases, and the other empty arm served as a blank control. Each arm is equipped with porous barrier to exclude any possible visual cues. The fruits were tested as non-sterilized and sterilized fruits. Chemical sterilization of fruits was done by dipping fruits in sodium hypochlorite solution (1%) for 1 min then rinsing with tap water and later with distilled water and then air-dried. Activated charcoal-filtered and humidified clean air

PheroFip 2019 106

was pumped uniformly through the arms at 20 ml min-1. Flies (n=10) were introduced at once to the stem of Y-tube and were observed for 1 h. Data were recorded based on the first choice made by a fly. The number of flies in fruit arm was expressed as a percentage of the total number of flies tested. Flies that did not make a choice were recorded as “no choice”. Flies were retrieved from olfactometer after 1 h, irrespective of the choice. After each test of 10 flies, the olfactometer was washed with distilled water and baked for 1 h at 150 °C. Each variety was tested separately in the olfactometer and each choice test was replicated 10 times thus the olfactory responses of 100 flies per variety were tested. The experiments were conducted in a laboratory at 23±1 °C, 65±5% relative humidity and 1000 lux light illuminance. Wilcoxon signed-rank test was used to determine whether there were differences in the attractiveness of sterilized fruits compared with non-sterilized fruits.

Results and discussion Both varieties of grapes tested were significantly more attractive than the control. It is found that D. suzukii female flies were more attracted to non-sterilized fruits of grapes more than sterilized fruits. Results depict that each variety showed significant difference in response of the flies for sterlized and non sterlized treatments- Vittoria var. (P <0.05) and Black magic var. (P <0.05). The highest attractiveness of fruits was recorded in white grapes var. Vittoria. The present study reports for the first time the behavioural response of D. suzukii to odors emitted by two varieties of grapes of agricultural importance in Italy. Our bioassays data indicate that olfaction has a major role in host selection for feeding and oviposition in grapes fruits. The attractiveness of female flies towards grapes varieties in the experiment reconfirms that D. suzukii is capable of infesting a wide range of hosts (Bellamy et al. 2013). However, flies attractiveness towards non-sterilized fruits can be related to microorganisms associated with fruits surface (Becher et al. 2012), therefore further studies are underway to identify microorganisms and volatiles emitted by them that elicit antennal and behavioural activity and may aid in the development of a selective and efficient synthetic lure. Such synthetic lure can be used as oviposition attractant rather than fermentation bait for effective monitoring and control of this pest.

Key words: Spotted wing drosophila, olfactometer, two-choice bioassays, grapes

Acknowledgements This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement no. 722642.

PheroFip 2019 107

Evidence for a marking pheromone attracting multiple oviposition in Drosophila suzukii

Gianfranco Anfora1,2*, Gabriella Tait2,3, M. Cristina Crava2,4, Daniel Dalton5, Silvia Carlin2, Vaughn M. Walton5, M. Valerio Rossi Stacconi5

1Center of Agriculture Food Environment (C3A), University of Trento, Via E. Mach 1, I-38100 San Michele all’Adige, Italy; 2Research and Innovation Centre, Fondazione Edmund Mach, Via E. Mach 1, I-38100 San Michele all’Adige, Trento, Italy; 3Department of Agricultural and Environmental Sciences, University of Udine, Italy; 4Department of Biology and Biotechnology, University of Pavia, Via Ferrata 9, I-27100 Pavia, Italy; 5Horticulture Department, Oregon State University, Corvallis, Oregon. *[email protected]

Abstract Drosophila suzukii Matsumura (Diptera: Drosophilidae) is an invasive fly native to Eastern Asia that has successfully invaded Europe and the Americas since its first detection in 2008 outside its native range. Its rapid spread and establishment is largely attributed to its ability to exploit for oviposition multiple intact host fruits ripening on plants; this ability is due to the presence of a sharp, enlarged ovipositor. However, the mechanisms of the oviposition site selection of D. suzukii are still matter of hypothesis. The aim of this work was therefore to elucidate behavioral and chemical aspects of short-range site selection of this species. Choice and non-choice oviposition tests indicated a preference towards egg-infested fruits, suggesting a role of the released liquid as a marking pheromone attracting multiple oviposition. Video recording evidenced several discrete and repeated steps during oviposition. In particular, the final step is the release of an anal secretion over the fruit surface near the oviposition site. Headspace extraction and gas chromatographic analysis revealed the presence of 17 compounds peculiar to the infested fruit. Further electrophysiological experiments indicated that both males and females D. suzukii antennae perceived six of these compounds. Finally, a synthetic blend composed by the six compounds in a ratio similar to that found in the post-oviposition secretion (marking liquid) increased the oviposition rate of conspecific females, suggesting that this secretion play a role in promoting oviposition and possibly aggregation in D. suzukii (marking pheromone). The knowledge gained with this study may accelerate establishment of control strategies based on the interference and disruption of the D. suzukii communication during the oviposition processes.

Key words: spotted wing drosophila, oviposition pheromone, integrated control

PheroFip 2019 108

Laboratory studies on potential oviposition deterrents for Drosophila suzukii

Christa Lethmayer1*, Matthias Wernicke1, Sylvia Blümel1

1AGES (Austrian Agency for Health and Food Safety), Institute for Sustainable Plant Production, Spargelfeldstraße 191, A-1220 Vienna, AUSTRIA *[email protected]

Abstract The spotted wing drosophila Drosophila suzukii (Drosophilidae, Diptera) was introduced to Europe in 2008 and established in almost every region of the continent since that time. This vinegar fly is very polyphagous and attacks ripening thin-skinned fruits of many cultivated fruit crops and wild host plants. Females lay their eggs with their serrated ovipositor into the fruits where the emerging larvae feed. Fruits become soft and rotten and therefore unmarketable. Severe damages and high economic losses are caused in fruit production. In the past decade many efforts to find efficient and sustainable integrated control strategies against D. suzukii were tested like sanitary measures, several trap types and attractants, different plant protection products and different varieties of exclusion options (netting). Another interesting possibility would be the potential use of repellent substances as an alternative method in the control of D. suzukii. However, only few investigations were dealing with repellent substances until now. Within the Interreg V-program/region Lake Constance (project: “Development of practicable measures against spotted winged drosophila to prevent economic damages”) new potentially repellent substances (different essential oils and substances from chemical groups like e.g. alcohol, ketone, ester, piperidine) against D. suzukii were tested in laboratory experiments in order to check their ability for reduction or prevention of its oviposition. In a first step two different “choice tests” (Dip-Test and modified DART-Test) were carried out with different kind of fruits (strawberries, cherries and raspberries) and laboratory reared D. suzukii from AGES. Subsequently also a “no choice-test” was carried out where only treated fruits (raspberries) were tested in a bioassay arena under controlled conditions. Evaluation of the tested substances is still under progress and results will be presented.

Keywords: Drosophila suzukii, control, oviposition, deterrent, repellent, laboratory test

PheroFip 2019 109

Spatial and temporal distribution of Drosphila suzukii in relation to fruit quality and coexistence with other arthropods, in sweet cherry orchards of British Columbia

Howard Thistlewood1,2*, Amanda Chamberlain1,2, Susanna Acheampong3

1Agriculture and Agri-Food Canada, Summerland Research and Development Centre, Summerland, British Columbia, V0H 1Z0, Canada; 2Department of Biology, University of British Columbia, Okanagan campus, 3187 University Way, Kelowna, BC, V1V 1V7; 3British Columbia Ministry of Agriculture, 200-1690 Powick Rd, Kelowna, BC, Canada V1X 7G5 *[email protected]

Abstract A vinegar fly, Drosophila suzukii Matsumura (Diptera: Drosophilidae), spotted wing drosophila, was first detected in British Columbia (B.C.) in 2009 (Thistlewood et al. 2012). Severe damage to soft fruit and berries from D. suzukii was reported, and its presence led to the increased use of insecticides on many crops. Studies commenced into this unknown invasive insect, as presented in overview in 2010 and 2014 (Thistlewood et al. 2012, 2016), from seasonal trapping on many crops and a spatio-temporal overwintering analysis (Thistlewood et al 2018), and concerning its ecology outside crops (Thistlewood et al 2019). We examine here the coexistence of D. suzukii, other drosophilids, and western cherry fruit fly, Rhagoletis indifferens, on the primary crop affected in our region, sweet cherry (Prunus avium), in the context of distribution on trees and individual cherries, and of fruit quality traits. For three years, collections were made from four cultivars ripening successively, in insecticide-free orchards. Twice weekly, mid-June to August, i.e. from unripe green fruit to mature over-ripe fruit, 128 cherries were picked from each cultivar (cv.), ‘Santina’, ‘Sonata’, ‘Lapins’ and ‘Staccato’ until post-harvest, for a total of 512 fruits. Subsamples were assayed using standard techniques for fruit quality traits of colour, firmness, soluble sugars, and titratable acids, and examined for insect presence under a microscope, or destructively (brown sugar test), or placed in rearing for three weeks. Additional collections of ground fruit, split versus non-split fruit, or until frost in autumn, were made in some years, and are discussed. For two seasons, R. indifferens was permitted to infest the fruit and attacked all cultivars simultaneously. By contrast, in two seasons with damaging population levels, D. suzukii infested each cultivar according to their successive ripening and harvest dates. The fruit quality results are presented in relation to the timing of entry of D. suzukii into fruit, for 8 cv. X year combinations. In a fourth season, we characterized the distribution of all drosophilids within the leaf canopy of five adjacent cultivars ripening successively in an orchard. For a seven week period, we regularly collected fruits from the canopies of trees (6-9 m high) as divided into four sectors (top or bottom thirds by north or south aspect) and from one central interior and two border (north, south) rows, for a total of circa 2,000 fruits. They were reared individually for two weeks, and drosophilid flies emerged from 72% of fruits, primarily D. suzukii, D. melanogaster, D. pseudoobscura, and D. hydei. Sap beetles (Coleoptera: Nitidulidae), cherry fruitworm Grapholita packardi (Lepidoptera: Tortricidae), and R. indifferens were also present. The results were analyzed using a fully-factorial design and showed that drosophilid oviposition is affected both by fruit phenology and microclimate. There was significant

PheroFip 2019 110

variation among cultivars; later ripening cultivars had a higher probability of being attacked than others. The mean number of drosophilids per canopy sector was significantly (P≤0.05 level) related to cultivar, row position, height, and aspect during early stages of infestation, and both cultivar and height were consistently significant during later stages of infestation. When populations were low (initial colonization), cherries collected from the interior rows and the (more shaded) north-facing row were significantly less likely to be infested than the south-facing row. Also, all drosophilids were more numerous in the bottom of the canopy but this effect was reduced when populations were high later in the season (secondary colonization). Interspecies facilitation occurred, whereby biological interactions within individual cherries influenced the distribution of ovipositing drosophilids, as identified using indices of aggregation to measure inter-specific and intra-specific interactions. The results suggest that oviposition by D. suzukii facilitates oviposition by saprophagous Drosophila species. In three of the four years, the earliest cv. (‘Santina’), reached harvest quality before fly attack, but not in the very warmest year, 2015. In all four years, the latest cv. (‘Staccato’) was attacked pre-harvest, but two cvs. with intermediate harvest dates were attacked to varying degrees pre-harvest. The findings provide improved understanding of the spatial and temporal dynamics of D. suzukii in an orchard crop, optimize the probability of detection and the efficacy of management programs, and add to the few studies that have quantified the temporal-spatial pattern of drosophilid oviposition in fruit. The results are discussed in the context of the monitoring and ecology of this invasive insect, and with implications for certain strategies of pest management.

Key words: Drosophila suzukii, sweet cherry, phenology, crop damage, fruit quality, distribution

Acknowledgements We thank B.C. Agriculture, Environment and Wildlife Fund, and B.C. Fruit Growers Association for support. Bradley Sinclair (Canadian Food Inspection Agency) verified the drosophilids.

References Thistlewood, H. M. A., Gill, P., Beers, E. H., Shearer, P. W., Walsh, D. B., Rozema, B. M., Acheampong, S., Castagnoli, S., Yee, W.L., Smytheman, P. & Whitener, A. 2018: Spatial Analysis of Seasonal Dynamics and Overwintering of Drosophila suzukii in the Okanagan-Columbia Basin, 2010–2014. Environ. Entomol. 47: 221-232. Thistlewood, H., Rozema, B. & Acheampong, S. 2016: Seasonal timing and infestation of sweet cherries and non-crop plants by Drosophila suzukii (Diptera: Drosophilidae) in and around fruit orchards of British Columbia, 2010-2013. IOBC-WPRS Bull. 112: 21-22. Thistlewood, H., Rozema, B. & Acheampong, S. 2018: Infestation and timing of use of non- crop plants by Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) in the Okanagan Basin, Canada. The Canadian Entomologist. In Press Thistlewood, H., Shearer, P. W., Steenwyk, B. V., Walton, V. & Acheampong, S. 2012: Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), a new pest of stone fruits in western North America. IOBC-WPRS Bull. 74: 133-137.

PheroFip 2019 111

Demonstration project “Exclusion netting for managing Spotted Wing Drosophila in fruit crops”

H. Vogt1*, B. Boehnke1**, K. Köppler2, C. Augel2, A. Wichura3, J. Lindstaedt4, J.-H. Wiebusch4, A. Engel5, S. Benz5, J. Saltzmann6, G. Eberhardt6

1Julius Kühn-Institut, Federal Research Centre for Cultivated Plants, Institute for Plant Protection in Fruit Crops and Viticulture, Schwabenheimer Str. 101, D-69221 Dossenheim; 2Landwirtschaftliches Technologiezentrum Augustenberg, Nesslerstraße 25, D-76227 Karlsruhe; 3Landwirtschaftskammer Niedersachsen, Pflanzenschutzamt, Wunstorfer Landstraße 9, D-30453 Hannover; 4Landwirtschaftskammer Niedersachsen, Moorende 53, D-21635 Jork; 5Landwirtschaftskammer Nordrhein-Westfalen, Pflanzenschutzdienst, Gartenstraße 11, D-50765 Köln-Auweiler; 6Julius Kühn- Institut, Federal Research Centre for Cultivated Plants, Institute for Strategies and Technology Assessment, Stahnsdorfer Damm 81, D-14532 Kleinmachnow, Germany *[email protected]; **[email protected]

Abstract The invasive Spotted Wing Drosophila Drosophila suzukii (Matsumura 1931) has become an enormous threat for European fruit cultivation. A technical possibility of control is the use of exclusion netting. In order to investigate and promote this technical approach in Germany, a demonstration project, funded by the German Federal Ministry of Food and Agriculture (BMEL) has started in 2017. It is a cooperation project with the Julius Kühn-Institut (JKI) Dossenheim as coordinator and the plant protection service of the participating Federal German states Baden-Württemberg, Niedersachsen and Nordrhein-Westfalen as partners. Up to now, the project includes 19 producers of cherry, raspberry, blackberry and blueberry. The farms had to apply for participation according to distinct criteria including that they had already installed nets. The tasks of JKI Dossenheim include the support of the plant protection services, the overall data evaluation, knowledge transfer and public relations of the project. The plant protection services support the demonstration farms in the implementation of the exclusion netting. Their project supervisors are in close contact with the growers and regularly collect data from the farms. Furthermore, the plant protection services also care for knowledge transfer, e.g. by initiating farm seminars in cooperation with the demonstration farms and the JKI. Another important part of the project is the analysis of the economic feasibility of already installed nets, which is covered by the JKI Institute for Strategies and Technology Assessment. For this purpose, investment and maintenance costs, labour input, crop yield and crop quality are assessed by interviews and investigations and then economically evaluated. Results from 2017 and 2018 will be presented with regard to the efficacy of the exclusion nets to control Spotted Wing Drosophila, the occurrence of other pests and beneficial insects, influence on the microclimate, technical problems, pollination deficiencies and the economic evaluation.

Key words: Exclusion netting, net, Drosophila suzukii, Spotted Wing Drosophila, economic analysis, microclimate

PheroFip 2019 112

Developing a push-pull system to manage Drosophila suzukii

Dong H. Cha1, Anna Wallingford2, Peter J. Landolt3, Gregory M. Loeb4

1USDA-ARS, US Pacific Basin Agricultural Research Center, Hilo, Hawaii, USA; 2University of New Hampshire Cooperative Extension, Durham, New Hampshire, USA; 3USDA-ARS, Yakima Agricultural Research Lab, Wapato, Washington, USA; 4Cornell University, Department of Entomology, Geneva, New York, USA

Abstract The spotted wing drosophila (Drosophila suzukii, SWD), introduced from Asia, is now a serious pest of numerous soft fruits throughout North America and Europe. Frequent use of insecticides is generally necessary to effectively manage SWD for susceptible berry crops. This is costly in terms of time, labor and materials as well as potentially for resistance to develop. It is also disruptive to established IPM programs. Hence, there is a great need for alternative control methods. We are particularly interested in identifying chemicals that are “attractive” and “repellent” to SWD and developing their use as a management tool. Initial screening of attractant and deterrent was focused on volatiles produced by different microbes. We developed and commercialized fermentation volatile based SWD attractant composed of acetic acid, ethanol, acetoin, and methionol (Cha et al. 2012, 2013, 2014, 2015, 2017, 2018). For deterrents, field assays found reduced SWD infestation of raspberries when combined with 1-octen-3-ol compared to control raspberries (Wallingford et al. 2016, 2017, 2018). Because 1-octen-3-ol requires relatively high concentrations, we are currently searching for novel repellents by investigating volatiles associated with the fruit pathogen Botrytis cinerea and by screening serendipitously discovered potential deterrents. Work continues on identifying the specific compounds responsible for the effect and their potential for management.

Key words: attractant, deterrent, push-pull, chemical ecology, spotted wing drosophila

References Cha, D. H., Adams, T., Rogg, H. & Landolt, P. J. 2012: Identification and field evaluation of fermentation volatiles from wine and vinegar that mediate attraction of spotted wing drosophila, Drosophila suzukii. Journal of Chemical Ecology 38: 1419-1431. Cha, D. H., Hesler, S. P., Cowles, R. S., Vogt, H., Loeb, G. M. & Landolt, P. J. 2013: Comparison of a synthetic chemical lure and standard fermented baits for trapping Drosophila suzukii (Diptera: Drosophilidae). Environmental Entomology 42: 1052-1060. Cha, D. H., Adams, T., Werle, C. T., Sampson, B. J., Adamczyk, J. J., Rogg, H. & Landolt, P. J. 2014: A four-component synthetic attractant for Drosophila suzukii (Diptera: Drosophilidae) isolated from fermented bait headspace. Pest Management Science 70: 324-331. Cha, D. H., Hesler, S. P., Park, S., Adams, T. B., Zack, R., Rogg, H., Loeb, G. M. & Landolt, P.J. 2015: Simpler is better: fewer nontarget insects trapped with a four-component chemical lure vs. a chemically more complex food-type bait for Drosophila suzukii. Entomologia Experimentalis Et Applicata 154: 251-260. Cha, D. H., Landolt, P. J. & Adams, T. B. 2017: Effect of chemical ratios of a microbial- based feeding attractant on trap catch of spotted wing drosophila (Diptera: Drosophilidae). Environmental Entomology 46: 907-915.

PheroFip 2019 113

Cha, D. H., Hesler, S. P., Wallingford, A. K., Zaman, F., Jentsch, P., Nyrop, J. & Leob, G. M. 2018: Comparison of commercial lures and food baits for early detection of fruit infestation risk by Drosophila suzukii (Diptera: Drosophilidae). Journal of Economic Entomology 111: 645-652. Wallingford, A. K., Hesler, S. P., Cha, D.H. & Loeb, G. M. 2016: Behavioral response of spotted-wing drosophila, Drosophila suzukii Matsumura, to aversive odors and a potential oviposition deterrent in the field. Pest Management Science 72: 701-706. Wallingford, A. K., Cha, D. H., Linn, C. E., Wolfin, M. S. & Loeb, G. M. 2017: Forum: Robust manipulations of pest insect behavior using repellents and practical application for integrated pest management. Environmental Entomology 46: 1041-1050. Wallingford, A. K., Cha, D. H. & Loeb, G. M. 2018: Evaluating a push-pull strategy for management of Drosophila suzukii Matsumura in red raspberry. Pest Management Science 74: 120-125.

PheroFip 2019 114

Exploring the potential of mass trapping Drosophila suzukii

Rik Clymans1,3,4*, Vincent Van Kerckvoorde1, Luk De Maeyer2, Tim Beliën1, Patrick De Clercq3, Dany Bylemans1,4

1pcfruit npo, Department of Zoology, Fruittuinweg 1, B-3800 Sint-Truiden, Belgium; 2Bayer AG, Crop Science Division, Jan Emiel Mommaertslaan 14, B-1831 Diegem, Belgium; 3Ghent University, Department of Plants and Crops, Coupure Links 653, B-9000 Gent, Belgium; 4KU Leuven, Department of Biosystems, Decroylaan 42, B-3001 Heverlee, Belgium *[email protected]

Abstract During the past decade, the Asian fruit fly Drosophila suzukii has become a key pest of all soft-skinned fruits in Europe and America. The current control relies on the intensive use of broad spectrum insecticides and hence forecloses the existing or developing IPM programs. Worldwide research is directed towards more sustainable control measures of D. suzukii like biological control (e.g. parasitoids), mechanical control (e.g. exclusion netting) and behavioural control (e.g. repellents, bait sprays and mass trapping). A great variety of attractants are developed/described for the detection and monitoring of D. suzukii. However, these attractants are generally considered unsatisfactory for a mass trapping strategy. This and the fact that experiments determining optimal trap density and efficacy of mass trapping can be complex as they demand large plots, could explain why little field research has been dedicated to the topic. Here we opted for a more fundamental approach to explore the potential of mass trapping D. suzukii for both standard fermentation baits and experimental chemical lures. A first method used, was developed by Suckling et al. (2015) and is based on trap interference in 4x4 trapping grids. In these experiments, the ratio of trap catch in outer traps to inner traps is an estimate of trap competition and thus overlap of attraction radii. A model for trap competition is used to know the level of overlap of attraction radii and therefore the potential of mass trapping at a certain inter-trap spacing. A second approach is the use of mark-release-recapture experiments with a grid of traps to determine parameters that can be used in a capture probability model. Hereto a method was developed to conveniently mark D. suzukii without affecting its survival and behaviour.

Key words: Diptera: Drosophilidae, semiochemicals, insect marking, integrated pest management, insect behaviour, stone and soft fruits

PheroFip 2019 115

An innovative management approach for spotted wing drosophila (Drosophila suzukii) using an environmentally friendly attract and kill formulation

Urban Spitaler1, Flavia Bianchi2, Irene Castellan3, Guillermo Rehermann4, Daniela Eisenstecken2, Paul G. Becher4, Sergio Angeli3, Silvia Schmidt1

1Division of Plant Protection, Research Centre Laimburg, South Tyrol, Ora, Italy 2Laboratory for Flavour and Metabolites, Research Centre Laimburg, South Tyrol, Ora, Italy 3Faculty of Science and Technology, Free University of Bozen-Bolzano, Bolzano, Italy 4Department of Plant Protection Biology, Swedish University of Agricultural Sciences, Alnarp, Sweden

Extended Abstract The spotted-wing drosophila, Drosophila suzukii (Matsumura) is a polyphagous invasive species in South Tyrol (Italy) which is responsible for major damages to local fruit farming and viticulture. The adult females attack soft skinned fruits of cultivated and spontaneous plants, especially berries, stone fruits and grapes. The fruit damage occurs shortly before harvest and is difficult to prevent with chemical agents. Stone fruit and berry cultivars represent a secondary source of income for many farmers in mountain areas of South Tyrol. Since the occurrence of D. suzukii the profitability of this production has been challenged and the chemical load on these cultures has greatly increased. During the last years several data related to the interactions between host plant-associated yeasts and D. suzukii were collected, and showed that some yeast strains favor larval development, female fecundity and stimulate the oviposition. These interactions are currently investigated by our research team. The main goal of this study is to develop an effective pest control strategy based on the behavioral manipulation of the insect to specific yeast lures, to reduce the load of chemical insecticide residues on the fruits. The expected result is an attract and kill formulation, which can attract D. suzukii females and males on target plant leaves surfaces. The project outputs include the identification of a lure with an attractive and a phagostimulant component and the elaboration of suitable application methods for different cultivation systems and cultural species. The lure should be combined with an insecticide and allow a reduction of the insecticide amount per ha. The methods should allow the transition from basic research to the field application of the final product. In this study, the following yeast species were tested: Saccharomyces cerevisiae, Saccharomycopsis vini, Candida sp., Issatchenkia terricola, Metschnikowia pulcherrima and three strains of Hanseniaspora uvarum. These yeast species were cultivated on two different culture media. To know to optimal harvesting time for all yeasts, the growth curves were determined. The yeasts were harvested at the beginning of the stationary phase. For every yeast culture the pH, temperature, biomass and OD was measured. At the defined harvesting time, an appropriate amount of the yeast culture was sampled for the further experiments.

PheroFip 2019 116

The attractant and feeding stimulant activity of the yeast cultures towards D. suzukii adults were determined in the laboratory and in the field and the most promising yeast species and substrate combination was defined. With a suitable yeast solution, an effective lure- combination was tested in the laboratory and in the field. Volatile compounds of yeast cultures were characterized by direct headspace analysis and gas chromatography-mass spectrometry (DHS-GC-MS). Semi-quantification was done for all compounds where authentic standards can be purchased. Altogether 16 compounds were characterized. Statistical analysis showed a higher abundance of ethyl acetate in Candida sp., while Saccharomycopsis vini was the only culture releasing a specific monoterpenoid compound. Saccharomyces cerevisiae released the highest rate of alcohols and ethyl esters, on the contrary, Hanseniaspora uvarum strains produced the lower amount of volatile compounds.The primary and secondary metabolites of all yeast species were characterized using mass spectrometry (LC-MS/MS) and flight mass spectrometry (LC- qTOF). Target analysis of extracellular compounds showed important differences in sugar and amino acids amounts between different yeast species, suggesting different consumptions of nutrients during fermentation. Volatile releases and metabolites analyses will be considered when choosing the most appropriate yeast species for the attract and kill formulation.

Key words: Drosophila suzukii, yeast, attractant, feeding stimulant, pest control strategy

Acknowledgements The project DROMYTAL is funded by “European Regional Development Fund (ERDF) 2014-2020“.

PheroFip 2019 117

Spotted wing drosophila: Extremely meteorosensitive - a base for the development of a Decision Support System

Kirsten Köppler1, Jeanette Jung2, Uwe Harzer3, Mandy Püffeld1, Marion Gradl3, Claudia Tebbe2, Alicia Winkler2, Paolo Raca2, Benno Kleinhenz2

1Center for Agricultural Technology Augustenberg (LTZ), Nesslerstr. 25, 76227 Karlsruhe, Germany 2Central Institute for Decision Support Systems in Crop Protection (ZEPP), Rüdesheimer Straße 60- 68, 55545 Bad Kreuznach, Germany, 3Agricultural Public Service Center Rheinpfalz, Breitenweg 71, 67435 Neustadt, Germany

Abstract The invasive Spotted Wing drosophila, Drosophila suzukii, is infesting and damaging stone and berry fruits and causing huge economical losses since serevral yaers in Germany. Population development and infestation level vary between different years, hence regulation measures are difficult to place in an optimal way. To develop a decision support system (DSS) for control measures, different factors influencing population development, activity and infestation of fruit crops is going to develop in a collaboration project. The DSS is expected to predict the population dynamics of D. suzukii and the actual pest infestation risk for the different crops, berries, stone fruits as well as grapes. For this resaon, the interaction of the entire life cycle and activity of D. suzukii with the most important meteorological factors, like temperature and humidity as well as landscape structures, wood, hedges or fields is described or functionally determined. Laboratory tests, monitoring activities and already published data will be combined within the DSS. In this presentation first focus is given on the influence of cold and freeszing temperatures during winter and landscape structure on population development will be shown. Second focus is given on the influence of temperature and humidity on flight and oviposition activity. As a first output of the DSS cherries will be assessed for their susceptibility depending on abiotic and ontogenetic factors.

Key words: Drosophila suzukii, Spotted wing drosophila, Decision Support System, climate, landscape structure

Acknowledgements This work is financially supported by the German Federal Ministry of Foof and Agriculture (BMEL) through the Federal Office for Agriculture and Food (BLE), grant number 2815HS013.

References Klick, J., Yang, W. Q., Walton, V. M., Dalton, D. T., Hagler, J. R., Dreves, A. J., Lee, J. C. & Bruck, D. J. 2016: Distribution and activity of Drosophila suzukii in cultivated raspberry and surrounding vegetation. Journal of Applied entomology 140(1-2): 37–46, DOI:10.1111/jen.12234. Pelton, E., Gratton, C., Isaacs, R., Timmeren, S. van, Blanton, A. & Guédot, C. 2016: Earlier activity of Drosophila suzukii in high woodland landscapes but relative abundance is unaffected. Journal of Pest Science 89(3): 725–733, DOI:10.1007/s10340-016-0733-z.

PheroFip 2019 118

Ryan, G. D., Emiljanowicz, L., Wilkinson, F., Kornya, M. & Newman, J.A. 2016: Thermal tolerances of the Spotted-Wing Drosophila Drosophila suzukii (Diptera: Drosophilidae). Journal of Economic Entomology, tow006.

PheroFip 2019 119

Explorative data analysis of a 7-year Drosophila suzukii monitoring program in Southwest Germany

Felix Briem1, Anto Raja Dominic², Burkhard Golla², Christoph Hoffmann³, Camilla Englert4, Annette Herz4, Heidrun Vogt1

1Julius Kühn-Institut, Institute for Plant Protection in Fruit Crops and Viticulture, Schwabenheimer Str. 011, D-69221 Dossenheim, Germany; ²Julius Kühn-Institut, Institute for Strategies and Technology Assessment, Stahnsdorfer Damm 81, D-14532 Kleinmachnow, Germany; ³Julius Kühn- Insitute, Institute for Plant Protection in Fruit Crops and Viticulture, Geilweilerhof, D-76833 Siebeldingen, Germany; 4Julius Kühn-Institute, Institute for Biological Control, Heinrichstraße 243, D-64287 Darmstadt, Germany

Abstract The invasive pest of soft-skinned fruits, Drosophila suzukii, was recorded for the first time in Germany in 2011. The same year, the Julius Kühn-Institut (JKI) set up a monitoring program to study the occurrence and activity of the flies in different habitats. Here we analyzed trap captures from 100 traps of the three participating JKI institutes for the effect of weather and immediate habitat on trap captures at different times of the year from late 2011 until early 2018. Those traps were distributed in semi-natural habitats, forested, agricultural and urban areas to represent a wide range of host plants or potential overwintering sites. We found that a stable population of D. suzukii was firmly established in southwest Germany after the mild winter of 2013/2014. We identified five phases in the annual population development of D. suzukii. Habitat types in the immediate vicinity of the trap and local weather conditions had a strong influence on trap captures varying with the season. Capture rates in forested areas were generally higher than those of orchards while traps in vineyards and urban areas had the lowest capture rates. Capture rates were negatively associated with the number of heat days and precipitation in late spring and summer while autumn, winter and early spring capture rates was negatively associated with the number of cold and ice days.

Key words: Drosophilidae; host plants; insect behaviour; insect traps; invasive species; monitoring system; population dynamics

References Briem, F, Dominic, A., Golla, B., Hoffmann, C., Englert, C., Herz, A. &Vogt, H. 2018: Explorative Data Analysis of Drosophila suzukii Trap Catches from a Seven-Year Monitoring Program in Southwest Germany. Insects 9(4)

PheroFip 2019 120

Three years of monitoring activities for invasive pest fruit flies in Austria

Alois Egartner1*, Christa Lethmayer1**, Richard A. Gottsberger1, Sylvia Blümel1

1Austrian Agency for Health and Food Safety (AGES), Institute for Sustainable Plant Production (NPP), Spargelfeldstr. 191, A-1220 Vienna, Austria *[email protected] **[email protected]

Abstract Fruit fly species (Diptera: Tephritidae) are among the most serious pests worldwide. While more than 4400 species are described, about 200 of them attack cultivated plants in orchards, vegetables or crops on arable land (Carroll et al. 2002). The number of native pest fruit fly species is low in Austria, however in regional monitoring activities also non-native fruit fly species (e.g. Rhagoletis cingulata, Ceratitis capitata) were observed in parts of Austria in the past (Egartner et al. 2010; Lethmayer, 2011). Furthermore, high numbers of tephritid interceptions at the entry points of the European Union (EC, 2017) encouraged the establishment of further monitoring activities for tephritids in Austria since 2016. Based on previous findings during Austrian monitoring activities, and because main host plants for important species of those genera are present in Austria, the monitoring focused on species from the genera Ceratitis and Bactrocera. Two traps (Maxitraps®, Tephritrap type) per site and genera were placed in selected peach orchards, on sites with alternative host plants, in the vicinity of market areas or at a potential point of entry. The number of monitoring sites was increased from 17 in 2016 to more than 30 in 2018, with the main focus on the city of Vienna. Mainly different parapheromones (Trimedlure, Methyl eugenol) to lure males were employed in the traps, and emptied in fortnightly intervals basically between May/June and September/October in 2016, 2017 and 2018. Identification of the caught tephritids was performed on morphological basis (White & Elson-Harris, 1992; Carroll et al. 2002; EPPO, 2011; EPPO, 2013) and confirmed with molecular diagnostic methods (including barcoding) for single flies (Van Houdt et al. 2010). In addition molecular sequencing was used to determine the potential source of some of the caught specimens. The fruit fly monitoring activities in Austria resulted in catches of several hundred fruit flies of Ceratitis capitata and single individuals of Bactrocera zonata and the Bactrocera dorsalis species complex. Detailed results of the different years will be presented.

Key words: fruit flies, Tephritidae, monitoring, Austria

References: Carroll, L. E., White, I. M., Freidberg, A., Norrbom, A. L., Dallwitz, M. J. & Thompson, F. C. 2002: Pest Fruit Flies of the World. Version: 13th September 2018. [WWW document] URL http://www.delta-intkey.com/ffa/index.htm. Cited 27 Sep. 2018. EC (European Commission - Directorate-General for Health and Food Safety) 2017: EUROPHYT-Interceptions - European Union Notification System for Plant Health Interceptions - Annual Report 2016. [WWW document] Ref. Ares(2017)3413447 - 06/07/2017. Cited 27 Sep. 2018. EPPO (European and Mediterranean Plant Protection Organization) 2011: Diagnostics - Ceratitis capitata. EPPO Bull. 41: 340-346.

PheroFip 2019 121

EPPO (European and Mediterranean Plant Protection Organization) 2013: Diagnostics - PM 7/114 (1) Bactrocera zonata. EPPO Bull. 43: 412-416. Egartner, A., Zeisner, Z., Hausdorf, H. & Blümel, S. 2010: First record of Rhagoletis cingulata (Loew) (Dipt., Tephritidae) in Austria. EPPO Bull. 40: 158-162. Lethmayer, C. 2011: Gefährliche Fliegen für Apfel und Co. Besseres Obst, 12: 4-5. Van Houdt, J. K., Breman, F. C., Virgilio, M. & DE Meyer, M. 2010: Recovering full DNA barcodes from natural history collections of Tephritid fruitflies (Tephritidae, Diptera) using mini barcodes. Mol Ecol Resour. 10(3): 459-65. White, I. M. & Elson-Harris, M. M. 1992: Fruit flies of economic significance: their identification and bionomics. C.A.B. International Paperback, Wallingford, Oxon, UK.

PheroFip 2019 122

Invasive tephritids (Diptera: Tephritidae) threatening European horticultural production and trade

Nikos T. Papadopoulos1*, Marc De Meyer2, David Nestel3, Slawomir Lux4, Darren J. Kriticos5, Stella Papanastasiou1, Kostas Zarpas1

1University of Thessaly, Department of Agriculture, Crop Production and Rural Environment, Laboratory of Entomology and Agricultural Zoology, Fytokou St., 38446 Volos, Greece; 2Royal Museum for Central Africa, Invertebrates Unit, Leuvensesteenweg 13, B3080 Tervuren, Belgium; 3Institute of Plant Protection, Dept. of Entomology, Volcani Ctr. ARO, Rishon Letzion, Israel; 4inSilico-IPM, 05-510 Ujejskiego 7, Konstancin-Jeziorna, Poland; 5CSIRO, GPO Box 1700, Canberra, ACT 2601, Australia *[email protected]

Abstract The European fruit industry, which provides an income for more than 1.5 million holdings, with more than €21billion of aggregated annual value, is under continuous threat from invasive alien pests entering the EU through trade, product movement and human mobility. Tropical fruit flies (Diptera: Tephritidae) are arguably the most important group of invasive pests commonly intercepted in European ports of entry (20-30% of all interceptions). Under current EU inspection regimes, newly introduced alien species may remain undetected for long periods, allowing populations to increase beyond levels that are economically or technically feasible to eradicate. Human movement facilitates random and scattered propagule dispersal. Expected increases in horticultural trade will escalate both the frequency of interceptions and the associated costs. Free movement of goods and people within the Schengen Zone of Europe, may further accelerate the spread of invasive fruit flies within the EU, and increases the difficulty in eradicating new invasions or slowing their spread. Out of more than 4,500 identified fruit fly species, Ceratitis capitata and Bactrocera dorsalis stand out as the most pestiferous and aggressive invaders, with frequent outbreaks in mainland USA, despite strict biosecurity regulations and procedures. Bactrocera spp. and Ceratitis spp. account for approximately 86% of all identified fruit fly species interceptions in Europe. Bactrocera dorsalis accounts for the highest propagule pressure (the most commonly intercepted pest) and has the capacity for remarkably rapid range expansion, e.g. soon after its first detection in 2003 in Kenya has now spread throughout sub-Saharan Africa and to adjacent islands. In Asia it is presently expanding its range into northern, more temperate areas of China, and it is detected annually in California. Likewise, Bactrocera zonata has invaded the north-eastern part of Africa and the Middle East, spreading southwards and eastwards; it is also detected repeatedly in California and is frequently intercepted in fruits imported into Europe. Ceratitis quilicii, (recently differentiated from C. rosa), although having a more limited geographical range might be better-adapted to low temperatures and therefore more suitable to be established in cooler European areas. Ceratitis capitata, although already widespread in the Mediterranean countries since early last century, appears to be expanding its range northwards. It is now frequently detected in Central Europe, previously thought to be too cold to allow population persistence. Coexistence of some of the above species (i.e. B. dorsalis, C. rosa, C. capitata in S. Africa, C. capitata and B. zonata in Egypt) imposes a huge burden to the biosecurity systems in Europe. Additional dipteran pest threats include the American species of the genus Anastrepha and Rhagoletis. This paper analyses the status of fruit fly invasion in Europe, explores its drivers including climate

PheroFip 2019 123

change and biological process, and provides a conceptual framework for developing specific response and mitigation actions aimed at preventing, detecting, eradicating and managing invasive fruit flies.

Key words: Oriental fruit fly, peach fruit fly, Mediterranean fruit fly, medfly, detection, interception, biosecurity, integrated pest management, Area Wide IPM

PheroFip 2019 124

SESSION 8

INTEGRATED PROTECTION OF FRUIT

CROPS

CHAIRPERSON: PETROS DAMOS

PheroFip 2019 125

A survey of integrated pest management in apple orchards in Norway

Bjørn Arild Hatteland1; Gaute Myren2, Iren Lunde Knutsen3, Kristin Kvamm-Lichtenfeld3, Endre Bjotveit3, Nina Trandem1

1Norwegian Institute for Bioeconomy Research, Biotechnology and Plant Health, Ås, Norway; 2Norwegian Agricultural Advisory Service, NLR Viken, Lier, Norway; 3Norwegian Agricultural Advisory Service, NLR Vest, Lofthus, Norway

Abstract Integrated pest management has been an important part of fruit production in Norway, especially during the 1980s and 1990s. Pesticide use was reduced due to forecasting systems and general knowledge of pests as well as monitoring methods. Currently, the focus on IPM has been renewed due to the implementation of the EU directive 2009/128/EC in 2015. In this study we investigated the IPM practices in Norwegian apple production. This was done by surveying 30 apple orchards during three consecutive years by the following methods: Insects and mites were collected by beat-tray samples prior to flowering and after flowering as well as by taking leaf samples after flowering. In addition, pheromone traps were applied for the codling moth (Cydia pomonella) and the fruitlet mining tortrix moth (Pammene rhediella). The apple harvest was analysed by counting and weighing apples from a random sample of trees in each orchard. A subsample of these apples was examined for potential damage, such as mechanical damage, frost damage, as well as damage by insect pests and fungal diseases. Finally, we monitored the measures taken by the farmers to manage pests, to analyse potential effects on yield, selected pests and beneficials. Damage by lepidopteran larvae was the most common insect fruit damage. However, only a few orchards had significant damage, which seemed to relate to high numbers of tortricid larvae early in the season. The most abundant pest insect sampled was the hawthorn psyllid (Cacopsylla melanoneura), which may cause damage during flowering. We found regional differences in the abundance of beneficial insects and mites. Anthocorid bugs were in relatively high numbers in orchards in western Norway, while lacewings (Chrysopidae) and predatory mites (Phytoseiidae) were more common in eastern Norway. We discuss our findings according to regional differences and farmer measures during the season.

PheroFip 2019 126

Effect of the apple cultivar on the duration of the development, survival and penetration rate of Grapholita molesta larval stage under field conditions

Carles Amat1,2, Cesca Alcala2, Dolors Bosch3, Jesús Avilla1, César Gemeno1, Lucía-Adriana Escudero-Colomar2*

1Department of Crop and Forest Sciences, University of Lleida (UdL), 25198 Lleida, Spain, 2Sustainable Plant Protection (Entomology), Mas Badia Agricultural Experimental Station, Institute for Agrifood Research and Technology (IRTA) 17134 La Tallada d’Empordá, Girona, Spain, 3Department of Sustainable Crop Protection, Institute for Agrifood Research and Technology (IRTA), 25198 Lleida, Spain * [email protected]

Abstract Apple (Malus × domestica Borkh) is usually considered a secondary host of Grapholita molesta Busck (Lepidoptera. Tortricidae). However, in different world areas, G. molesta has been increasable found in apple orchards causing major damage, as for example in the Girona region (NE extreme of Spain). During the growing season, G. molesta affects different apple cultivars which may affect differently the OFM fitness. The aim of this study is to determine the development time, survivorship and penetration rate of G. molesta larvae in three different apple cultivars (“Gala”, “Golden” and “Fuji”) under field conditions. Two wild populations were used; one from Girona province only developed on apple and another from Lleida province where it can develop on apple and peach (Prunus persica Stokes). The study has been conducted in commercial orchards of each variety without insecticides treatments. When each cultivar reached 75% of fruit ripening, 300 24-h old eggs of each population were individually placed on an apple, and the fruits were insulated with mesh sleeves. Fruits were daily checked to register larvae penetration in fruit as well as the presence of pupae in the corrugated cardboard pieces placed inside the sleeves to facilitate pupation. All apples were collected after 100 degree days had passed since the first larva cocoon had been detected, registering in laboratory the number of penetrations, dead and living larvae as well as some fruit quality parameters. During the experiments in the three varieties, two data loggers registered the temperature and HR in each orchard. Results showed that the development time, the survivorship and the penetration rate of G. molesta was significantly affected by the apple variety, with Gala being the least suitable for the species. There were no significant differences in their responses to these parameters between the two populations studied. The implication of these results in the management of the pest is discussed.

Key words: Grapholita molesta, apple, development time, survivorship

PheroFip 2019 127

Long term trials to reduce pesticide use in fruits- results of EUFRUIT project partners

Barbara Egger1, Andreas Naef1*, Christelle Lacroix2, Hinrich Holthusen3, Jordi Cabrefiga4, Franziska Zavagli5

1Agroscope, Wädenswil, Switzerland; 2INRA, Montfavet Cedex, France; 3Esteburg, Jork, Germany, 4IRTA, Girona, Spain; 5CTIFL, Prigonrieux, France *[email protected]

Abstract Fruits have to be protected against a series of pests and diseases, which is impossible with a single technique. Different approaches such as prophylactic measures, beneficial insects, mechanical intervention, biocontrol products, and less susceptible varieties are used. However, pesticides still play an important role in fruit production since quality requirements of the market are increasing and the risk for significant economic losses is high. This is in contradiction to consumer expectations for residue-free fruits and minimal pesticide use. Within the EUFRUIT project, several partners have established long term trials with combinations of alternative plant protection techniques to compare the agronomic performance and the economic feasibility of these strategies to actual growers’ practices. The BioREco orchard (2005 - 2015) was the first system experiment in fruit tree production in France that developed such an approach and addressed the control of the major pests and diseases of apple, i.e. scab, codling moth, and powdery mildew. Three systems (Conventional, CON; Low Input, LI; Organic, ORG) were planted with three apple cultivars differing in disease susceptibility (Smoothee, a Golden Delicious type cultivar susceptible to scab; Ariane, a scab-resistant cultivar; and Melrose, a low-susceptibility cultivar). Cultivation practices (cultivar choice, tree training, supervised fertilization and irrigation), and natural enemy-mediated processes (selective pesticide use and reduction of the mowing frequency) were considered. Direct measures included sanitation practices and alternative methods to synthetic pesticides, such as mechanical weeding, mating disruption, microbiological and biological control with granulosis virus and entomopathogenic nematodes. Lastly, applications were based on decision support systems such as forecasts of meteorological conditions, observations in orchards, and prediction models (scab, codling moth). Compared to the regional reference, it was possible to reduce the use of pesticides by an average of 38 to 45% with the LI and ORG system. This reduction was achieved for similar yield levels in LI and CON systems. The yield was lower in the ORG system where fruit damage may be higher. The abundance and diversity of the studied biological communities (earthworms, earwigs, spiders) varied depending on the system and year. More generally, the environmental impact of the systems was reduced by decreasing the use of pesticides. Production costs were higher in LI and ORG than in CON systems, but without price premium in LI. Also in France, a national network on pesticide reduction in apple orchards started in 2012 within the ECOPHYTO national program. The idea was to evaluate 27 systems located in six different regions during six years. On each location, the low input systems are compared to an orchard representing the conventional practices. On average between 2013 and 2017 the low input strategy achieved a reduction of the “global treatment frequency indicator” (TFI) as follows: – max. 40% reduction with a scab sensitive variety, where some treatments during the first and second scab contamination periods could be avoided, combined with mating disruption and biocontrol agent insecticides (low codling moth pressure) and adapted treatments against aphids. – more than 58% reduction with a scab

PheroFip 2019 128

sensitive variety, by using a rain cover against apple scab combined with mating disruption and different types of insecticides (chemicals and biocontrol agent) or combined with an exclusion net and limited complementary insecticides against codling moth (low pressure). The same level of reduction was also obtained by adapting the doses of all products to the trees’ volume and stage. In each case, the protection against aphids was adapted to their pressure. – between 55 and 65% reduction with a scab resistant variety, where the main apple scab protection had been covered by natural products combined with exclusion nets or/and mating disruption depending on the codling moths pressure. – More than 75% reduction with a scab resistant variety produced in an organic production system or by adapting doses with conventional products, both combined with exclusion nets or mating disruption. In Northern Germany, regional programs on pesticides and pesticide residual reduction in apple started in 2009. Five different plant protection systems were evaluated in one orchard over five years. The focus was on preventing detectable pesticide residues at the time of harvest. The average number of detectable residuals could be reduced to 1.4 when application was completely stopped or switched to pesticides authorized for organic production only after blossom, compared to 3.3 detectable pesticide residues when synthetic pesticides were applied until harvest. Fruit losses were high after 5 month of cold storage when no pesticides were applied after blossom (more than 40% due to storage diseases). Using fungicides authorized in organic production after blossom reduced fruit losses by more than 60% while synthetic fungicides reduced losses by up to 70%. In Switzerland, during a nine-year period, Agroscope tested a crop protection strategy for apples with the objective to produce fruits without residues. This Low Residue (LR) strategy was compared to the Integrated Production (IP) and the Organic Production (OP). Attributes of the economic sustainability have been evaluated in order to compare all three strategies. With the chosen plant protection, fertilizer and thinning programs in the LR strategy, it was possible, even with susceptible varieties such as Golden Delicious, to reach a yield comparable to the IP-strategy. However, the pack out for Golden Delicious was about 10% lower and for Topaz even 20% lower in the LR strategy compared to the IP strategy. All varieties showed significant losses due to storage diseases (mainly Gloeosporium rot) in the LR and the OP strategy. The evaluation showed that the lower pack out for the LR strategy adversely affects the profitability. Unlike for the OP strategy, there is no premium price to compensate the lower yield and the higher production risk of the low LR-strategy. Results showed that by choosing an adapted cultivar it should be possible to increase the pack out after storage and decrease pack out loss with positive consequences on the income variability. In Catalonia, since 2011, a cooperative project between the Agriculture Department of this region, IRTA and the fruit sector, FRUIT.NET, is running to optimize the use of pesticides and minimize the residues in apples, pears, peaches and citrus. In the case of apple production, the project is based on predictive models for apple scab (apple), on the priority of use of alternative methods (mating disruption, mass trapping) to chemicals and on avoiding the longer remaining active ingredient treatments close to harvest. The results of the program are promising: in apples the use of insecticides was decreased by up to 35% and the use of fungicides was reduced by 24%, achieving a low residues level on fruits at harvest (less than 20% of MLR). Long term trials as described above are continuing to implement new methods. These trials will lead to practical recommendations for growers and demonstrate eco-friendly fruit production systems to retailers and consumers.

Key words: Iow input fruit production, pesticide residues, codling moth, apple scab, storage diseases

PheroFip 2019 129

Putting IPM in practice in apple: an 8-year Case Study from Southern Sweden

Marco Tasin1, Sanja Manduric2, Weronika Swiergiel1, Joakim Pålsson1, Birgitta Rämert1, Mario Porcel1,3

1SLU, Department of Plant Protection Biology, 230 53 Alnarp, Sweden; 2Swedish Board of Agriculture, Sweden; 3Colombian Corporation for Agricultural Research (Corpoica), C. I. La Libertad, Vía Puerto López km 17, Meta, Colombia

Abstract We assessed here the co-innovation process that was carried out in Southern Sweden during 2010-2017 by researchers and apple production actors. We aimed at the development of sustainable IPM methods that, with the help of an agroecological whole systems approach, would be both desirable and feasible to use in practice by apple growers. Whilst a novel pest management strategy based on sex-pheromone was rather promptly appreciated, the enhancement of natural enemies through flower strips took a much longer learning. The knowledge gap due to the reduced number of extension advisors and the divergence between the cost incurred when implementing low-impact pest control methods and reduced profitability of apple were pointed out as major contradictions by the stakeholders. We argue here that a reinforced regional agroecological scheme support along with the expansion of public advisory service would reduce the farmer economic risk and share the responsibility for a safer environment and healthier fruits. In connection to that, local authorities should be provided with resources and competence to allow for safety assessments of candidate low-risk plant protection products at the regional scale. As a conclusion, we recognized that in the Skåne region sustainable apple orchard management through feasible and desirable plant protection strategies could not be achieved exclusively by focussing on the efficiency of the tools because the cost-effectiveness and thus the implementation of such tools depended greatly on the simultaneous co-innovation of the socio-technical system. Stakeholders need to reflect upon their vision to propose long term environmentally sound ideas providing a base to promote, finance and adopt safe plant protection strategies within the Skåne region.

Key words: agroecological approach, biological control, functional diversity, Integrated Pest Management, participatory approach.

PheroFip 2019 130

Temperature driven stochastic distribution models and simulation of codling moth phenology

Petros Damos1, Polyxeni Soulopoulou2, Thomas Thomidis2

1Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece; 2Alexander Technological and Educational Institute of Thessaloniki, Department of Agricultural Technology

Extended Abstract Phenological models have been important tools used to improve the management of codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae) in many apple producing areas worldwide (Damos et al. 2015). In this work studies were conducted in the prefecture of Imathia in Northern Greece to test a stage specific phenology model that has been developed ad hoc for predicting the emergence of important phenological events of codling moth, Cydia pomonella at field conditions. Conceptually, the current modeling framework has been proposed by Osawa et al. (1983), to describe bud phenology of balsam fir [Abies balsamea (L.) Mill.] and later modified by Dennis et al. (1986) to insect development using a logistic distribution to simplify calculations. The model is a departure compared to most empirical based phenology models since it is of stochastic nature and is based on a set of logistic probability distribution functions (pdf) that evolve in time. Each pdf express the proportion of the population which has completed a particular phenological event and/or developmental stage αi at time t, where i corresponds to one of the following stages: oviposition, egg hatch, larval development, pupal development and adult emergence. Stage succession occurs when the amount of development passed through a stage specific developmental threshold H at t, which is a measure of the physiological time interval expressed in degree-days. Thus, for each stage, the proportion of the population pi in developmental stage i at sampling time t is given by the logistic probability density function (pdf) with the general condition: 푝푖(푡) = Pr⁡[푋(푡) = 푖], while the probability that a given individual is at a specific developmental stage at time t=H is:⁡푝푖(푡) = Pr[푋(푡) > 퐻], where H is the stage specific thermal constant expressed in degree-days (DD). Thus, the general form of the model consist of a set of logistic equations, each of which gives the cumulative probabilities pi(t) of individuals residing in stages αi(t)={1,2…,n}. As time, t, evolves the joint probabilities of 훮 individuals reside at least in one of the n stages are: ∑1 푝푖,푡 = 1. The parameters of the model correspond to the central moments of the pdf and are the stage specific developmental threshold, A and the scale parameter λ which express the variability. These parameters were estimated using data from laboratory growth studies conducted at constant temperature and solved using the the maximum likelihood estimate procedure obtained as the solution of the following maximization problem (Matlab 2000):⁡휃̂ = arg max휃퐶훩 ln⁡[퐿(휃; 휉)]. Where Θ is the parameter space, ξ are the observed data (the sample),⁡퐿(휃; 휉) is the likelihood of the sample, which depends on the parameter θ; and arg max is the operator which gives the parameter for which the log-likelihood ln[퐿(휃; 휉)] takes its maximum value. In all cases the degree days for the emergence and distribution of each developmental event were estimated using a linear model of heat summation: 퐷(푇 − 푇퐿) = 퐻 (Damos and Savopoulou-Soultanin 2012) where D, T, TL, and H, represent the duration of development (days), environmental (mean/isothermal) temperature (oC), the estimated developmental zero

PheroFip 2019 131

(lower developmental threshold) temperature, and the effective cumulative temperature or thermal constant, respectively. According to the model projections, there is a very high probability, p=0.999, of observing adults of the first flight generation until 333DD but a very low probability of finding adults of the second flight generation. Furthermore, at 472DD the probability of observing individuals belonging to the first flight generation practically diminishes, p=0.05, but there is a very high probability of observing moths that belong to the second flight generation, p=0.948. At 1175DD, the probability of observing individuals that belong to the second flight generation is very low, p=0.003, while there is a long period in which the probability of observing moths that belong either to the second or the third generation is zero. In particular, at 575DD the probability of finding an individual to lay eggs is p=0.15. However, there is nearly the same probability of egg hatch, p=0.36, and larval completion p=0.313, while at the same time the probability of pupal completion is very low, p=0.001. Thus, there is a critical period between 450~650DD in which a significant overlapping of developmental events occurs, including moth population decline, peaks in oviposition and egg hatch, as well as the emergence of the first larvae. Concluding, codling moth management in Greece is characterized as an intensive program of repeated insecticide sprays based on accurate timing of stage selective biorational pesticides to protect the fruit from larval injuries. The current probalistic degree-day model provides a means of accurately predicting C. pomonella biological events. However, to be applied on a regular basis, following-up of the model prediction should be further evaluated for other biological events and such as egg laying period and egg hatch.

Key words: Cydia pomonella, forecasting model, IPM

References Damos, P., Savopoulou-Soultani, M. 2012: Temperature-Driven Models for Insect Development and Vital Thermal Requirements, Psyche Volume 2012, Article ID 123405, 13 pages. Damos, P., Escudero Colomar, L. E. & Ioriatti, C. 2015: Integrated fruit production and pest management in Europe: The apple case study and how far we are from the original concept? Insects 6(3): 626–657. Dennis, B., Kemp, W. P. & Beckwith, R. C. 1986: Stochastic model of insect phenology: Estimation and testing. Environmental Entomology 15: 540-546. MATLAB 6.1, The MathWorks Inc., Natick, MA, (2000). Osawa, A., Shoemaker, C. A. & Stedinger, J. R. 1983: A stochastic model of balsam fir bud phenology utilizing maximum likelihood parameter estimation. Forest Science 29: 478- 490.

PheroFip 2019 132

Towards a knowledge-based decision support system for integrated control of woolly apple aphid, Eriosoma lanigerum, with maximal biological suppression by the parasitoid Apheninus mali

Eva Bangels1, Ammar Alhmedi1, Dany Bylemans1, Tim Belien1

1pcfruit, Zoology Department, Fruittuinweg 1, B-3800 Sint-Truiden, Belgium

Abstract The woolly apple aphid Eriosoma lanigerum (Homoptera: Aphidiae) is an important pest in apple orchards worldwide. Since the withdrawal or restricted use of certain broad-spectrum insecticides, E. lanigerum has become one of the most severe pests in apple growing areas across Western Europe. Their protective wax coating makes them particularly difficult to control. At present, effective limitation of woolly aphid populations relies on a good synergy between chemical control treatments and biological suppression by beneficial arthropods, especially by its main specific natural enemy the parasitoid Apheninus mali (Hymenoptera: Apelinidae). Optimal control strategies take advantage of a thorough understanding of the life cycle of E. lanigerum, with a good knowledge of the timing of their migration waves, as well as the activity of A. mali and its parasitation activity. In order to develop a knowledge-based decision support system detailed monitoring data of both species were collected in the field (region of Sint-Truiden, Belgium) for a period of eight years (2010-2018). Aphelinus mali flights were monitored in the field, starting before flowering until at minimum the end of the second generation flight. The seasonal occurrence of the most important management stages of E. lanigerum, e.g. start of wool production or activity in spring and migration of crawlers from colonies towards flower clusters or shoots, were monitored. All obtained data were compared with historical and literature data and analysed in a population dynamics phenological model. The model was used to generate predictions of the seasonal development of E. lanigerum and the biological control activity by A. mali. Based on the outcomes and interpretations, thorough decision support was generated for the optimal timing and positioning of control treatments in order to achieve effective control of E. lanigerum with maximal biological suppression by its main natural enemy. This was tested and evaluated in field trials.

Key words: woolly apple aphid Eriosoma lanigerum, parasitoid Apheninus mali, population dynamics, decision support

PheroFip 2019 133

Plum pocket caused by Taphrina pruni can also be problematic in commercial plum growing

Jorunn Børve1, Arne Stensvand1,2

1Norwegian Institute of Bioeconomy Research, P.O. Box 115, 1431 Ås, Norway; 2Norwegian University of Life Sciences, Universitetstunet 3, 1433 Ås, Norway

Abstract Plum pocket caused by Taprina pruni is known as a home garden disease of European plum (Prunus domestica), because it usually appears in cultivars of minor importance in commercial fruit production and on unsprayed trees. Over the last decade, the incidence of plum pocket has increased in commercial plum orchards in Norway, especially in cv. Mallard. In 2016, 2017 and 2018 experiments with timing of fungicide applications were performed in order to find optimal time of treatment to avoid the disease. Copper oxide (Nordox 75WG) or dithianon (Delan WG) were applied at different times from the autumn of 2015 and until prior to flowering in 2018. Number of fruit with plum pocket were counted, and in some of the plots, yields were also documented. Results varied among seasons and plots, but in general copper applications reduced the incidence of plum pocket, and the application of copper should be performed before green tissue was visible on the buds. Dithianon alone had minor effect and in combination with copper it resulted in no additional effect. The results are discussed in relation to climatic conditions and phenology of the plum trees.

Key words: bud development, cultivars, precipitation

PheroFip 2019 134

Pseudomonas syringae on young sweet cherry trees in Norway

Jorunn Børve1, Juliana Irina Spies Perminow1, Marcel Wenneker2, May Bente Brurberg1,3, Arne Stensvand1,3

1Norwegian Institute of Bioeconomy Research, P.O. Box 115, 1431 Ås, Norway; 2Wageningen University & Research, Lingewal 1, 6668 LA Randwijk, The Netherlands; 3Norwegian University of Life Sciences, Universitetstunet 3, 1433 Ås, Norway

Abstract Bacterial canker caused by different strains (pathovars, pv.) of Pseudomonas syringae, is a common disease in sweet cherry (Prunus avium) cultivation. Sweet cherry trees may have either visible or latent infections of P. syringae at the time of planting. Reduced yield and in some cases tree death are the consequences of the infections. Because of the cool, wet climate in Norway, the disease is of major economical importance. Nearly all sweet cherry trees planted in Norway are imported from nurseries in the Netherlands, Belgium or Austria. In 2015-2018, disease development was followed after planting of sweet cherry trees that had naturally occurring infections. Disease development varied from orchard to orchard and among cultivars. A high number of trees displaying symptoms of bacterial infections ranging from low to high severity were observed. The most commonly observed symptoms in the planting year were discolored areas and cankers at the heading back point, or in defeathering wounds between this point and the graft union. In the following years, blighted fruit spurs, dead branches, oozing cankers, or dead trees were observed. The attack differed among orchards. In one orchard with four cultivars (Giorgia, Lapins, TechloVan and Van), planted in 2015, 0 to 30% of the trees from each cultivar were with visible symptoms in the planting year, however, by the autumn of 2017 85 to 100% of the trees had symptoms of bacterial canker. The cultivars Giorgia and TechloVan in another orchard had 32 or 9% trees with symptoms after planting in 2015 and in 2018 55 and 91%, respectively. In a nursery study over two years, trees were assessed for symptoms at lifting (harvest) and after the cold store period prior to planting. The incidence of visible symptoms of bacterial canker increased from none at lifting to 33 and 46% after cold storage on cv. Giorgia in the two years, respectively. From the cold stored trees P. syringae pv. syringae and in one case P. syringae pv. morsprunorum were isolated. Growers are now advised to carefully look for bacterial cankers before and after planting and remove infected trees, avoid sub-optimal planting locations, optimize watering and fertilizing to avoid stressing the trees during and after planting, and to protect them against further infections by preventive applications with a copper fungicide.

Key words: inoculum, Prunus avium

PheroFip 2019 135

POSTER PRESENTATIONS ABSTRACTS

PheroFip 2019 136

SESSION 1

MANAGING BENEFICIAL INSECTS BY

SEMIOCHEMICALS

CHAIRPERSON: ANDREA LUCCHI

PheroFip 2019 137

Perspectives of multi-species lures for attracting Chrysopidae

Sándor Koczor1, Ferenc Szentkirályi1, Miklós Tóth1

1Plant Protection Institute, CAR, HAS; H-1022 Budapest, Herman Ottó u. 15., Hungary

Abstract Green lacewings (Chrysopidae) are important predators of several soft-bodied insect pests, with preference for Sternorrhyncha, especially aphids. The Chrysoperla carnea species complex is of special importance in agroecosystems. A ternary floral bait was developed which is a powerful attractant to both sexes of these lacewings, furthermore females were found to lay their eggs in the vicinity of the baits, which is of crucial importance in respect of biological control. Nevertheless adults of Chrysoperla spp. are not predatory, thus it would be beneficial to apply stimuli, which may be attractive to a wider range of green lacewings, including predatory adults of Chrysopa spp. In order to find potential compounds for development of multi-species lures, field experiments were conducted in Hungary. In the presentation we are aiming to give an overview of these results. In our field experiments, an aphid sex pheromone compound, nepetalactol attracted males of Chrysopa species, predominantly C. formosa, a species present in agroecosystems, however in combination with the ternary floral bait, the number of attracted Chrysoperla spp. decreased markedly. On the other hand squalene was also found to be attractive to C. formosa. Interestingly this compound was also attractive to males only. When tested in combination with the ternary floral bait, no significant interaction was found. Our results indicate that the combination of the ternary floral bait and squalene could be promising for simultaneous attraction of Chrysoperla spp. and males of Chrysopa spp. Perspectives, challenges and potential practical applications will be discussed.

Acknowledgements The current research was partially supported by the National Research, Development and Innovation Office NKFIH - PD115938 grant and the János Bolyai Research Scholarship of the Hungarian Academy of Sciences.

PheroFip 2019 138

SESSION 2

NEW DEVELOPMENTS IN PEST

MONITORING METHODS

CHAIRPERSON: TIM BELIEN

PheroFip 2019 139

Attractant for the invasive mealybug citrus pest, Delottococcus aberiae (Hemiptera: Pseudococcidae)

Sandra Vacas1, Ismael Navarro2, Javier Marzo2, Vicente Navarro-Llopis1, Jaime Primo1

1CEQA-Instituto Agroforestal del Mediterráneo, Universitat Politècnica de València, Camino de Vera s/n, edificio 6C-5ª planta, 46022 Valencia (Valencia), Spain; 2Ecología y Protección Agrícola SL, Pol. Ind. Ciutat de Carlet, 46240 Carlet (Valencia), Spain

Abstract Native to sub-Saharan Africa, Delottococcus aberiae (De Lotto) (Hemiptera: Pseudococcidae) was commonly found in South Africa on wild olive trees and flowering shrubs but irregularly distributed in citrus orchards of the north of the country. However, this mealybug has been recently reported as causing severe damage to citrus in Eastern Spain, posing a serious threat to the rest of the Mediterranean area and many other regions through international trade of citrus plants or fruits. Detection, surveillance and population monitoring are essential tools to control invasive pests but little is still known about D. aberiae biology, behavior and attractants. Once provided evidence of male attraction to virgin females, this study dealt with the isolation and identification of the attractant emitted by D. aberiae virgin females to effectively lure conspecific males. The volatile profile of D. aberiae virgin and mated females was studied by aeration and collection of effluvia in adsorbent matrix, to detect virgin female specific compounds. The collected volatiles were retrieved by solvent extraction and the resulting extracts were subjected to gas chromatography coupled to mass spectrometry (GC-MS). Analyses revealed a candidate compound which was isolated by gravity column and preparative gas chromatography. Molecular structure was elucidated according to GC-MS and nuclear magnetic resonance spectroscopy data. Although absolute configuration has not yet been determined, a racemic mixture of isomers was synthesized to confirm its structure and perform activity tests.

Key words: attractant; lure; sex pheromone; mealybug; Pseudococcidae; volatile

PheroFip 2019 140

Attracting Lobesia botrana with microbial volatiles: effect of dose and blend ratio

Sebastian Larsson Herrera1, Peter Rikk1, Gabriella Köblös, Teun Dekker1, Bela Peter Molnar2, Marco Tasin1

1SLU, Department of Plant Protection Biology, 230 53 Alnarp, Sweden; 2Zoology Department, Plant Protection Institute, Centre for Agricultural Research, Hungarian Academy of Sciences, Budapest, Hungary

Abstract Although the technology of mating disruption is available for L. botrana since almost three decades, implementation is only achieved on a restricted number of viticultural districts in the world. Factors that limited the spread of this environmentally friendly technology are among others, the higher labour required to hand apply the dispenser in comparison to standard practices based on pesticide, the challenge to involved a critical number of stakeholders to reach an area wide approach and the lack of reliable attractant to monitor pest populations within a pheromone permeated crop. Volatile attractants for both female and male insect can be used as a monitoring tool with a potential to importantly decrease the use of synthetic pesticides within both conventional and pheromone programs. Rather than using host-plant volatiles, which readily are masked by the main crop, we tested here the potential of using volatiles that signify microbial breakdown, to assure that the volatiles stand out against the background odour. To test this hypothesis, we investigated the behavioural response of the grapevine moth to synthetic microbial compounds in vineyards. Transparent delta traps baited with a two-component blend of acetic acid and 2-phenyl ethanol caught significant numbers of both sexes in comparison with unbaited traps. Only a few moths were caught by either of the two components alone. Although moths were attracted to a various ratio of volatiles in this blend, the minimum amount necessary to elicit attraction was compound dependent. With a view on practical applications, additional field experiments are on-progress to improve both the attraction and the selectivity of the identified blends.

Key words: grapevine moth, integrated pest management, kairomone, trapping, monitoring

PheroFip 2019 141

EAG responses of Lobesia botranamales and females collected from Vitis vinifera and Daphne gnidium to host-plant volatiles and sex pheromone compounds

Alicia Pérez-Aparicio1, Luis Miguel Torres-Vila2, César Gemeno1

1University of Lleida, Department of Crop and Forest Sciences, Av. Alcalde Rovira Roure 191, 25198 Lleida, Spain 2Servicio de Sanidad Vegetal, Consejería de Medio Ambiente y Rural PAyT, Junta de Extremadura, Avda. LuisRamallo s/n, 06800 Mérida, Badajoz, Spain.Servicio

Abstract We analyzed electroantennogram (EAG) responses of male and female adults of Lobesia botrana (Denis et Schiffermüller) (Lepidoptera: Tortricidae) collected as larvae from the host plants Vitis vinifera L.and Daphne gnidium L. The host-plant compounds tested were either V. vinifera-specific [1-octen-3-ol, (E)-β-farnesene, (E)-4,8-Dimethyl-1,3,7-nonatriene], D. gnidium-specific [2-ethyl-hexan-1-ol, benzothiazole, linalool-oxide, ethyl benzanoate], or were shared by both host-plants (linalool, methyl salicylate). Sex pheromone compounds were also tested. Male response to the major pheromone component (E7, Z9-12:Ac) was higher than to any other stimuli, whereas response to the minor pheromone compounds (E7, Z9-12:OH, Z9-12:Ac) did not differ from the response to the plant odorants. Female response to the pheromone was lower or not significantly different than to some plant odorants. Response to pheromone compounds was higher in males than in females whereas response to plant volatiles was not different between sexes, except for methyl salicylate, which elicited higher response in females. (E)-β-farnesene elicited higher EAG responses than several other plant odorants in both sexes. No significant interaction between host-plant odorant and sex indicated lack of sex specialization for host-plant volatile detection. Larval host-plant affected overall EAG responses (higher amplitude in V. vinifera), however, lack of significant interaction between plant volatile and larval host-plant suggested lack of specialization for plant-volatile detection between V. vinifera and D. gnidium individuals. Our study provides a first assessment of host differences in olfaction, and the lack of host-odor specificity suggests that if an evolutionary host-shift from D. gnidium to V. vinifera has taken place in the past it has not been accompanied by a significant modification of the sensory system. Reproductive isolation, if any, has not modified sex pheromone detection either.

Key words: electroantennogram, host plant, Lobesia botrana, sex pheromone, Tortricidae, volatiles

PheroFip 2019 142

Observations on flight activity and voltinism of codling moth Cydia pomonella L. in western central part of Latvia in 2016-2018.

Edite Jakobsone1*, Laura Ozolina-Pole1

1Latvian Plant Protection Research Centre Ltd., Struktoru iela 14a, LV-1039, Riga, Latvia; *[email protected]

Abstract A key apple pest codling moth Cydia pomonella L.has been considered univoltine in Latvian conditions, with only rare cases of incomplete second generation. In the light of recent advancements on the topic of climate change, a shift towards bivoltinism has to be considered. Change of voltinism would also require a shift in recommendations for codling moth management. We carried out observations on flight activity of codling moth, using traps baited with sex pheromone lures. Observations were carried out in one orchard for three consecutive years and various voltinism patterns were observed ranging from typical univoltine development in 2016 to a pattern strongly suggesting bivoltine development in 2018. It suggests enough developmental plasticity in codling moth to adjust its voltinism to changing environment. It stays unclear if the second generation is complete.

Introduction Codling moth Cydia pomonella is a key pest in apple orchards in Latvia (Priedītis 1996), but nowadays the access to decision support systems and modern insecticides allows Latvian farmers to achieve suficient control. So far codling moth has been considered univoltine in Latvia (Priedītis 1996). However, according to to the global climate change predictions, a prognosis was made by Stoeckli et. al. in 2012 that a global shift in codling moth voltinism might take place. If the shift occours, significant adjustments in codling moth management reccomendations have to be made.

Material and methods Observation site: Observations were carried out in an apple orchard located in western central part of Latvia, 57°01'58.1"N 22°55'03.8"E. The orchard was managed using principles of integrated pest management. Meteorological data was obtained from hydrometeorological station Stende, located 28 km North-West from the observation site. Trapping methods: Each year four transparent delta traps with sticky inserts and four green top funnel traps were placed in a lattice pattern in the orchard. The traps were baited with codling moth female mating pheromone lures following the directions of manufacturer Pherobank BV. Codling moth males caught were counted aproximately weekly. Exposure began from 10th to 25th of May, and ended from 12th to 30th of August.

Results and discussion Codling moth flight activity: In 2016 the main flight took place until 27.06.16. The number of males caught during main flight responded to fluctuations in temperature, a brief period of temperatures lower than the long term average from 05.06.16 to 14.06.16 was folloved by temporary lower trap catches. It matches with the opinions of Ozols (1973) and Priedītis (1996) who state that flight activity is possible only when temperature exceeds 15°C. Overall the flight pattern showed typical univoltine development.

PheroFip 2019 143

In 2017 active flight period was extended, ranging from 02.06.17 to 03.08.17, with individual males still getting caught until 17.08.17. According to Ozols (1973) usually the flight activity dissipates in first tertile of July. Partialy the extended flight period in our observations can be attributed to improvements in detection, however our observations suggest existance of some larvae that pupated around first or second tertile of July and hatched in August. Flight pattern in 2017 showed a possibility of a part of population developing a second generation. In future direct examination for pupae should be carried out in July, to confirm its development. The vegetation season of 2018 was distinguished by unusually warm and dry weather conditions. Codling moth male trap catches showed two distinct peaks. The first peak took place from 17.05.18 until 21.06.18, reaching its maximum on 24.05.18. The second peak was observed from 10.08.18 until 22.08.18. This pattern suggests a strong evidence that large part of the population experienced bivoltine development in 2018. However it is unclear if the second generation will successfully develop until maturity, as the development from oviposition to mature larva lasts from 35 to 47 days (Priedītis 1996), and many of the apple varieties grown in this particular orchard are harvested earlier. Implications for farmers: These observations imply that if the climat change trend towards warmer summers continues, bivoltine development can become more common. If that turns out to be the case, adjustments in reccomendations of codling moth control should be made.

Key words: Malus x domestica, monitoring, pheromones

Acknowledgements We thank Rural Support Service for financing this research.

References Ozols, E. 1973: Lauksaimniecības entomoloģija, Rīga, izdevniecība "Zvaigzne", 405-409 Priedītis, A. 1996: Kultūraugu kaitēkļi, Rīga, apgāds "Zvaigzne ABC", 224-227 Stoeckli, S., Hirschi, M., Spirig, C., Calanca, P., Rotach, M. W. & Samietz, J. 2012: Impact of Climate Change on Voltinism and Prospective Diapause Induction of a Global Pest Insect – Cydia pomonella (L.), https://doi.org/10.1371/journal.pone.0035723

PheroFip 2019 144

Improving the cost efficiency and practice implementation of automated camera-based monitoring of tortricid pests (Cydia pomonella) in pome fruit orchards

Ammar Alhmedi1, Tim Belien1, Bart De Ketelaere2, Dany Bylemans1

1 pcfruit, Zoology Department, Sint-Truiden, Belgium 2Katholieke Univ Leuven, Div Mech Biostat & Sensors MeBioS, Leuven, Belgium

Abstract The current applied practice of manual monitoring in fruit orchards has some major drawbacks. It is a labor intensive practice and therefore costly, due to the need of weekly visual inspections of spatially (widely) distributed traps. It is also error-prone, as it relies on visual judgment of (often non-expert) personnel. Because of these limitations the manually collected data typically are heterogeneous and sparse in terms of geographic and temporal coverage, making them unsuitable for accurately mapping of pest population dynamics. Therefore there is an increasing interest in electronic devices that can automatically detect/identify pests and can simultaneously monitor a large number of sites on a daily basis without requiring human presence. In the camera-based automatic monitoring approach digital images of trapped target pest(s) are automatically captured and sent to a server. The pests are then identified and counted either manually by human experts looking at them on the screen, or automatically by image-recognition software, which can gradually improve by machine learning methods (so called artificial intelligence or deep learning methods). Although automatic image-recognition technology has advanced dramatically in recent years, and an increasing number of studies are published demonstrating successes in machine image- based classification and even identification of pest species, this method is not widely applied in fruit growing practice yet. One of the main reasons for this is the relatively high cost per camera-trap device. In order to make this much more cost efficient (and thus economically achievable) we investigated the possibilities to monitor simultaneously several target pests with one and the same camera-trap device in pome fruit orchards. For this purpose, we evaluated the efficacy of monitoring traps in which different pheromones for different tortricid pests are combined. Firstly, we developed laboratory olfactometer choice tests to screen the behavioural responses of codling moth (Cydia pomonella) and other fruit tortricid pests to different pheromone combinations. In these olfactory assays we demonstrated that Cydia pomonella males are able to detect the correct pheromone even if it was combined with other moth pheromones. Furthermore, we validated the efficacy of camera-equipped pheromone traps in the field.

Key words: automatic monitoring, camera traps, Cydia pomonella, pheromone combinations, olfactometer choice tests

PheroFip 2019 145

Failure of commercial sex-pheromone monitoring lures in the assessment of codling moth presence: an emerging problem in South Tyrol

Silvia Schmidt1, Peter Neulichedl1, Josef Österreicher2, Paul Pernter2, Michele Preti3, Sergio Angeli3

1Laimburg Research Centre, Institute for Plant Health, 39040 Ora, Italy; 2South Tyrolean Advisory Service for Fruit- and Winegrowing, 39011 Lana, Italy, 3Faculty of Science and Technology, Free University of Bozen-Bolzano, Bolzano, Italy

Abstract In South Tyrol (North Italy), the apple growing area represents about 18 000 ha. Since 1992 mating disruption started to be applied as control method against codling moth reaching already 25% of the area in 1996 and about 90% of the total apple growing area in 2015. This control technique allowed to reduce the number of insecticide applications against Cydia pomonella L., which however remains the key insect pest species for apple growers of the region. Therefore, attention had still to be paid to monitor failures of mating disruption techniques. For that purposes usually high loaded sex-pheromone (superlure 12 mg, Pherobank, The Netherlands) baited monitoring traps are recommended by the South Tyrolean advisory service to avoid failures of mating disruption and increase cloud/plume cover signalled by cumulative males captures. Starting from 2006, an experimental apple orchard of cv. Golden Delicious) was established at Laimburg Research Centre where neither chemical control nor mating disruption have been applied against C. pomonella, to be used as control plot to assess the population dynamics of the species. Traps baited with monitoring codlemone-lures (Pherobank ) were checked yearly from April until September two times a week for males captures and weekly fruit damage control were performed in order to define important phenological phases, like starting of first generation flight, first fruit penetration as well as peak of fruit penetration. These are useful information also with regard to time of insecticide applications. During the last 3 years we observed failures in the assessment of codling moth population and in damage control by using both, the monitoring codlemone-lure in our control orchard and also the superlure traps in several apple orchards under mating disruption of South Tyrol. In this situation, periodical fruit damage controls have to be performed also in mated disrupted apple orchards to prevent unexpected fruit damages and optimize application timing of insecticide applications. In 2017, in order to ameliorate the use of the sex pheromone trap as tool for IPM strategies for codling moth, we decided to perform a comparative study using different codlemone loaded lures of 1mg, 2 mg, 4mg and 8 mg (Pherobank) in two untreated control plots with homogeneous infestation of codling moth. The archived results show important differences in flight activity curves suggesting that the initial load of codlemone to therubber septa plays a significant role for an efficient trapping method. Companies should carefully evaluate changes in load or rather adapt the load for different climatic or local situations. Direct head space analyses of the used septa were performed in GC-MS with a representative number of septa belonging to two different commercial companies and to different production batches. Gas-chromatographic analyses were also carried out after a period of field exposure of the septa. First examinations confirmed the reliability of the chosen method to test the releasing rate of each septum

PheroFip 2019 146

Population dynamic and flight activity will be further assessed in 2019 in three untreated plots of South Tyrol with selected rubber septa and the data will be analyzed in comparison with of head space results. This investigation should reveal if production charges differs significantly with one another. If case the results of these study will not explain the capture failure in codling moth, we shall take into account the possibility of an ecological adaptation of the species to the codlemone, the main component of pheromone plume, by South Tyrolean codling moth populations, perhaps due to a twenty-year old exposure to mating disruption.

Key words: Cydia pomonella, product quality control, codlemone baited traps, IPM tool, head space analysis

PheroFip 2019 147

Use of phenylacetonitrile plus acetic acid as a multi-species trapping method to monitor the leafrollers Pandemis spp. and Choristoneura rosaceana along with Cydia pomonella (Lepidoptera: Tortricidae) in apple

Esteban Basoalto1, Alan L. Knight2, Gary J. R. Judd3

1Instituto de Producción y Sanidad Vegetal, Facultad de Ciencias Agrarias, Universidad Austral de Chile, Campus Isla Teja, Valdivia 5090000, Chile. [email protected] 2Yakima Agricultural Research Laboratory, USDA-ARS, 5230 Konnowac Pass Rd, Wapato, Washington 98951, USA. [email protected] 3Agriculture and Agri-Food Canada, Summerland Research and Development Centre, Box 5000, 4200 Highway 97, Summerland, British Columbia, Canada V0H1Z0. [email protected]

Abstract In the western United States, Pandemis pyrusana (Kearfott), Pandemis limitata (Robinson), and Choristoneura rosaceana Harris, are the key leafroller pest species attacking apple. Insecticides often are used to manage these pests, although sex pheromone mating disruption has been developed and implemented on a relatively small acreage. Sex pheromone-baited traps are used to monitor adult populations to assess density and phenology of these pests, but pest managers feel these traps are insensitive to local population densities within the orchards. However, more recent studies have identified that the combination of acetic acid with either phenylacetonitrile or 2-phenylethanol produced a very effective bisexual lure for these species (Knight et al. 2017; Giacomuzzi et al., 2017; Judd et al. 2017; Basoalto et al. 2017). Adoption of these new binary lures may improve our ability to monitor leafroller populations in orchards treated with sex pheromones for mating disruption. Another useful attribute of these lures is their attraction to female moths. The sex ratio of moths caught in baited traps has been close to 1:1 in most studies. Codling moth, Cydia pomonella (L.), is the key tortricid pest of pome fruit orchards, and growers use a greater density of traps for this species than other tortricid leafrollers. Some efforts to reduce monitoring costs have previously considered combining sex pheromone lures of C. pomonella and leafrollers in a single trap (Knight et al. 2014). Several studies have already examined the possible interaction of these new attractants with sex pheromones and other plant volatiles. For example, traps baited with leafroller sex pheromone caught significantly fewer males when lures loaded with either 2-phenylethanol or phenylacetonitrile alone (C. rosaceana) or acetic acid plus 2-phenylethanol (P. limitata) were added. The addition of (E)-4,8-dimethyl-1,3,7-nonatriene to traps to enhance the catch of C. pomonella has previously been reported (Knight et al. 2015) and may be most useful in orchards treated with mating disruption technologies incorporating pear ester. However, co- monitoring C. pomonella and C. rosaceana in traps including (E)-4,8-dimethyl-1,3,7- nonatriene significantly reduced the catch of latter species (Knight et al. 2017). Interestingly, traps baited with phenylacetonitrile plus acetic acid caught significantly more males, females, and total moths than the number of males caught in traps baited with a commercial sex pheromone lure. The evaporation rate of the acetic acid co-lure was shown to be an important factor affecting catches of P. pyrusana with phenylacetonitrile. Adding phenylacetonitrile to traps baited with pear ester, and acetic acid significantly reduced both total and female moth catches of codling moth. However, neither C. pomonella nor P. pyrusana catch were impacted when phenylacetonitrile and acetic acid were added to traps baited with pear ester plus the sex pheromone of codling moth (Basoalto et al. 2017). In aggregate, these studies suggest that the optimal development of a multi-species trapping approach combining sex

PheroFip 2019 148

pheromones with plant and food volatiles will likely require extensive testing of blend combinations and perhaps trade-offs in efficacy.

Key words: kairomones; host plant volatiles; codling moth; leafroller; trapping; sex pheromone

References Basoalto, E., Hilton, R., Judd, G. J. R., Knight, A. L. El-Sayed, A. M. & Suckling, D.M. 2017: Evaluating the use of phenylacetonitrile plus acetic acid to monitor Pandemis pyrusana and Cydia pomonella (Lepidoptera: Tortricidae) in apple. Fla. Entomol. 100: 761-766. Giacomuzzi, V., Mattheis, J. P., Basoalto, E., Angeli, S. & Knight, A. L. 2017: Survey of conspecific herbivore-induced volatiles from apple as possible attractants for Pandemis pyrusana (Lepidoptera: Tortricidae) Pest. Manage. Sci. 73: 1837-1845. Judd, G. J. R., Knight, A. L. & El-Sayed A. M. 2017: Trapping Pandemis limitata (Lepidoptera: Tortricidae) moths with mixtures of acetic acid, caterpillar-induced apple- leaf volatiles, and sex pheromone. The Cana. Entomol. 149: 813-822. Knight, A. L., Hilton, R., Basoalto, E. & Stelinski, L. L. 2014: Use of glacial acetic acid to enhance bisexual monitoring of tortricid pests with kairomone lures in pome fruits. Environ. Entomol. 43: 1628-1640. Knight, A. L., Basoalto, E., Katalin, J. & El-Sayed, A. M. 2015: A binary host plant volatile lure combined with acetic acid to monitor codling moth (Lepidoptera: Tortricidae). Environ. Entomol. 44: 1434-1440. Knight, A. L., El-Sayed, A. M., Judd, G. J. R. & Basoalto, E. 2017: Development of 2- phenylethanol plus acetic acid lures to monitor obliquebanded leafroller (Lepidoptera: Tortricidae) under mating disruption. J. Appl. Entomol. 141: 729-739.

PheroFip 2019 149

Improved monitoring of oriental fruit moth (Lepidoptera: Tortricidae) with terpinyl acetate plus acetic acid membrane lures in Uruguayan orchards

María Valentina Mujica1, Alan L. Knight2

1INIA, Instituto Nacional de Investigación Agropecuaria,Estación experimental Las Brujas, Ruta 48 km 10 Canelones 90200, Uruguay. [email protected]; 2USDA-ARS, Yakima Agricultural Research Laboratory, Konnowac Pass Rd. Wapato, Washington 98951, USA. [email protected]

Abstract The use of sex pheromones for the management of oriental fruit moth, Grapholita molesta (Busck), has been broadly adopted throughout its worldwide distribution in both stone (Prunus spp.) and pome (Malus domestica Borkhausen, and Pyrus spp.) fruits and now includes the use of hand-applied dispensers, aerosols, sprayables and attract and kill formulations. One limitation with sex pheromone-based management programs is the difficulty in monitoring pest populations with sex pheromone-baited traps. A series of studies have recently reported on the use of terpinyl acetate in a sticky trap (Cichon et al. 2013; Knight et al. 2013) and the use of a new pheromone lure that uses the combination for the three-component sex pheromone of G. molesta plus the sex pheromone of Cydia pomonella (L.) (Knight et al. 2014; Knight et al. 2016). Recent studies with other important tortricids, such as C. pomonella (Knight et al. 2015), and several tortricid leafrollers (Knight et al. 2017; Basoalto et al. 2017) have shown catches can be significantly increased when kairomones are combined with acetic acid. Thus, studies with G. molesta in Uruguay were part of a world- wide study to test several combinations of terpinyl acetate, acetic acid, and several kairomones in delta traps with sticky liners (Mujica et al. 2018). Male and female moth catches of G. molesta in traps baited with various septa and membrane lures were evaluated in stone and pome fruit orchards treated with sex pheromones for mating disruption from 2015 to 2017. Membrane lures were loaded with terpinyl acetate (TA), acetic acid (AA) and (Z)-3- hexenyl acetate alone or in combinations. Two septa lures were loaded with either the three- component sex pheromone blend for G. molesta alone or in combination with codlemone (2- PH), the sex pheromone of C. pomonella. A third septum lure included the combination sex pheromone blend plus pear ester, (E,Z)-2,4-ethyl decadienoate (2-PH/PE), and a fourth septum was loaded with only β-ocimene. The first study was conducted from 15 December to 18 January (2015 – 2016) in a peach orchard treated with Isomate OFM dispensers (Pacific Biocontrol) at 250/ha and was adjacent to commercial apple orchards. This study compared five lures, including OFM-PH, 2-PH, 2-PH + BO, 2-PH + TA + AA and 2-PH + TA + AA + BO. The second study was conducted during 2016-2017 and consisted of three trials. Trial 1 was conducted from 25 January to 10 February in a “Fuji” apple block treated with OFM/CM puffers (Suterra) at 2.5 units/ha. The second trial was conducted from 26 January to 16 February in peaches treated with Checkmate OFM (Suterra) at 250 dispensers per ha. The third study was conducted from 26 January to 16 February in peaches treated with the paste formulation, SPLAT OFM (ISCA Technologies) at 3.7 kg/ha. Results were consistent across studies and showed that the addition of β-ocimene or (Z)-3- hexenyl acetate did not increase moth catches. The addition of pear ester to the sex pheromone lure only marginally increased moth catches. The use of TA and AA together significantly increased moth catches compared with the use of only one of the two components. The addition of TA/AA significantly increased moth catches when combined with the 2-PH lure. The TA/AA lure also allowed traps to catch both sexes. Catch of C. pomonella with the 2-PH lure was comparable to the

PheroFip 2019 150

use of codlemone. Optimization of these complex lures can likely further improve managers ability to monitor G. molesta and help to develop multispecies tortricid lures for use in individual traps. The complexity of combining sex pheromones with host plant, and microbial volatiles for a suite of species may be too great to create one optimized lure. Instead, the use of suboptimal lures with some sufficient attractiveness across all species might be sufficient to establish action thresholds and to reduce monitoring costs.

Key words: apple, Cydia pomonella, Grapholita molesta, mating disruption, peach

Acknowledgements We would all like to thank Bill Lingren, Trécé Inc., Adair, OK, for providing the experimental septa and membrane lures used extensively in these trials.

References Basoalto, E., Hilton, R., Judd, G. J. R., Knight, A. L. El-Sayed, A. M. et al. 2017: Evaluating the use of phenylacetonitrile plus acetic acid to monitor Pandemis pyrusana and Cydia pomonella (Lepidoptera: Tortricidae) in apple. Fla. Entomol. 100: 761-766. Cichon, L., Fuentes-Contreras, E., Garrido, S., Lago, J., Barros-Parada, W. et al. 2013: Monitoring oriental fruit moth (Lepidoptera: Tortricidae) with sticky traps baited with terpinyl acetate and sex pheromone. J. Appl. Entomol. 137: 275-281. Knight, A., Basoalto, E., Hilton, R., Molinari, F, Zoller, B. et al. 2013: Monitoring oriental fruit moth (Lepidoptera: Tortricide) with the Ajar bait trap in orchards under mating disruption. J. Appl. Entomol. 137: 650-660. Knight, A., Cichon, L., Lago, J, Fuentes-Contreras, E, Barros-Parada, W., et al. 2014: Monitoring oriental fruit moth and codling moth (Lepidoptera: Tortricidae) with combinations of pheromones and kairomones. J. Appl. Entomol., 138: 783-794. Knight, A. L., Basoalto, E., Katalin, J. & El-Sayed, A. M. 2015: A binary host plant volatile lure combined with acetic acid to monitor codling moth (Lepidoptera: Tortricidae). Environ. Entomol. 44: 1434-1440. Knight, A. L., Basoalto, E. & Stelinski, L. L. 2016: Variability in the efficacy of sex pheromone lures for monitoring oriental fruit moth (Lepidoptera: Tortricidae). J. Appl. Entomol. 140: 261-267. Knight, A. L., El-Sayed, A. M., Judd, G. J. R. & Basoalto, E. 2017: Development of 2- phenylethanol plus acetic acid lures to monitor obliquebanded leafroller (Lepidoptera: Tortricidae) under mating disruption. J. Appl. Entomol. 141: 729-739. Mujica, V., Preti, M., Basoalto, E., Cichon, L. Fuentes-Contreras, E. et al. 2018: Improved monitoring of oriental fruit moth (Lepidoptera: Tortricidae) with terpinyl acetate plus acetic acid membrane lures. J. Appl. Entomol. 142: 731-744.

PheroFip 2019 151

The determination of the effectiveness of pheromone traps for the control Box Tree Moth - Cydalima perspectalis in Georgia

Archil Supatashvili1, Medea Burjanadze1, Giorgi Mamadashvili2, Natia Iordanishvili2, Beqa Berdzenishvili3, Temel Gorkturk 4

1Agricultural University of Georgia, 240 David Agmashenebeli Alley, 0159 Tbilisi, Georgia; 2 LEPL National Forestry Agency, Ministry of Environment Protection and Agriculture of Georgia, 0114 Gulua st.6, Tbilisi, Georgia; 3The National Botanical Garden of Georgia, Botanikuri st,Tbilisi, Georgia; 4 Artvin Çoruh University, Forest Faculty, Artvin, Turkey [email protected]

Abstract The Box Tree Moth (BTM) - Cydalima perspectalis (Walker, 1859) (Lepidoptera; Crambidae) was introduce in Georgia and in the next year it began to defoliate Buxus spp in large quantities. Today the situation is quite alarming in Western Georgia, that BTM damage Buxus colchica, which is endemic species of Caucasian flora and threatened by habitat loss. The larvae feed on leaves and shoots, caused serious damages, defoliating box trees, causing economic, social and environment problems. 2017-2018, WitaTrap® Funnel trap system, with pheromone CYDAWIT® (Witasek, Pflanzenschutz, GmbH, Austria), were installed for the monitoring and control of BTM. A long term trapping was conducted at two location of boxwood forest in Tsageri - Ambrolauri (South slop of Grate Caucasian mountain range) in West Georgia. 450 pheromone trap were set out on the 150 ha at least ten day before the pest is expected to emerge and at the proper height above the ground or in the plant canopy. Three trap per ha were placed, where prevailing winds was carry the pheromone into the forest area. Attracted moth by the pheromone fall into cap-ture container and cannot fly out. The pheromone traps were emptied and new dispensers were added two time at the during the flying period of C.perspectalis. The number of captured adults were variated from 11 to 176 moth from one trap. In total approximately 93000 (2017) and 74000 (2018) of C.perspectalis captured in Tsageri – Ambrolauri region.

Key word: Cydalima perspectalis, pheromone trap, Buxus, monitoring

PheroFip 2019 152

Comparison of counting methods for apple leafcurling midge on pheromone traps

Daniel Cormier1, Franz Vanoosthuyse1, Gérald Chouinard1

1Research and Development Institute for the Agri-Environment (IRDA), 335 rang des Vingt-Cinq Est, Saint-Bruno-de-Montarville, J3V 0G7 (Quebec) Canada

Abstract When scouting for apple leafcurling midge adults, Dasineura mali (Kieffer) (Diptera: Cecidomyiidae), observers may deal with counts from as low as a few specimens up to a few thousands on the sticky Delta traps baited with sex pheromone used to monitor this insect. The project aimed to evaluate if scouts can adequately count the number of specimens captured using a 10x magnifying glass. It also aimed to check whether it is necessary to count the specimens on the entire sticky liner or if a part of it suffices. Using the following attributes: number of antennal articles, pattern of wing venation and presence of haemolymph spots on the tibiae, an inexperienced observer counted adults caught on 41 sticky traps, first using a 10x magnifying glass and then, a 6.4x binocular microscope. Afterwards, traps were counted again, this time by an experienced observer using the binocular microscope. Numbers from both observer types were then compared. Estimations of adult densities with the 10x magnifying glass were also performed by the same inexperienced observer; adults from 25 (every fourth square) and 50 % (every other square) of the sticky liner surface were counted. The results indicate that the inexperienced observer did as well as the experienced one with both the magnifying glass and the binocular microscope, except for traps containing low numbers of adults. False identifications were made for both low and high catches and were more frequent with the 10x magnifier. As expected, counting adults on reduced trapping surfaces was not representative for traps with few specimens due to the aggregated pattern of catches. Overall, the 10x magnifier was appropriate for counting the number of D. mali adults caught on pheromone sticky traps. Traps should be fully counted when less than 50 adults are present, whereas observing 50 % of the surface, as opposed to 25 %, should provide good estimates when less than 400 adults are present.

Key words: density estimation, apple pest, integrated pest management

PheroFip 2019 153

Progress towards identification of a pheromone of the asparagus beetle, Crioceris asparagi (Coleoptera; Chrysomelidae)

Steven Harte1*, Daniel Bray1, Sam Brown2, Jude Bennison2, David Hall1

1Natural Resources Institute, University of Greenwich, Chatham, ME4 4TB, UK; 2Agricultural Development and Advisory Service, ADAS Boxworth, Cambridgeshire, CB23 4NN *[email protected]

Abstract The asparagus beetle, Crioceris asparagi L. (Coleoptera; Chrysomelidae) has become a widespread pest of asparagus, Asparagus officinalis (asparagus). There are no known attractants for C. asparagi and much of its biology is unknown. Volatiles were collected from individual beetles and a candidate pheromone component was detected in collections from some individuals of the first generation but not the second. This compound elicited an electroantennographic response from C. asparagi and the mass spectrum indicated it is a 19- carbon hydrocarbon with four double bonds. Complete structural elucidation and synthesis are in progress.

Introduction The asparagus beetle, Crioceris asparagi L. (Coleoptera; Chrysomelidae) originated in the Mediterranean region but now occurs wherever asparagus, Asparagus officinalis, is grown. Asparagus appears to be the sole source of food with infestations causing severe damage to both the foliage and the commercially valuable spears. Currently, there are no known attractants for C. asparagi and much of its biology is still unknown, although a male- produced aggregation pheromone has been identified in the closely-related cereal leaf beetle, Oulema melanopus L. (Cossé et al 2002). An attractive pheromone for C. asparagi could be used in population monitoring and possibly control, and contribute towards a greater understanding of its biology.

Material and methods Insects: Crioceris asparagi beetles were collected as adults and as late larvae or pupae from asparagus fields in Cambridge and Kent. Larvae and pupae were reared through to adults in individual containers in the laboratory at ADAS. Collections were made of beetles of the second generation in June-July 2017 and the overwintering generation in May-June 2018. Beetles were sexed by dissection and examination of the genitalia. Collection and analysis of volatiles: Volatiles were collected from individual beetles on Porapak resin and eluted with dichloromethane. Collections were analysed by gas chromatography (GC) coupled to mass spectrometry (MS) on both polar (DBWax) and non- polar (VF5) columns. Collections were also analysed by GC coupled to electroantennographic (EAG) recording from a beetle antenna with intermittent, “puffed” delivery of the column effluent to the EAG preparation.

Results and discussion A peak was observed in GC-MS analyses of 18 out of 66 collections from C. asaparagi beetles of the overwintering generation which elicited an EAG response from a beetle antenna (Figure 1), indicating it was a potential pheromone component. This compound was not detected in 30 collections from the second generation, adding further support to it being a

PheroFip 2019 154

component of a sex or aggregation pheromone since only the overwintering beetles mate and those of the second generation overwinter as adults.

Figure 1: GC-EAG analysis of collection of volatiles from Crioceris asparagi (lower trace GC-FID, upper EAG response to intermittent delivery of column effluent; * EAG response).

The sex was determined of most of the beetles that produced this compond in significant quantities. The compound was apparently produced by both sexes (8:7 males:females). This was surprising as pheromones are almost exclusively emitted by one sex for any given species. The mass spectrum had a molecular ion at m/z 260 and base peak at m/z 79 (Figure 2). The GC retention indices were consistent with it being a hydrocarbon with 19 carbon atoms and four non-conjugated double bonds. Hydrogenation produced nonadecane, confirming that it is a 19-carbon, straight chain hydrocarbon. Authentic (Z,Z,Z)-1,3,6,9- and (Z,Z,Z,Z)- 3,6,9,12-nonadecatetraenes had similar but not identical mass spectra and retention times. This structure is very different from the hydroxyketone identified from O. melanopus L. by Cossé et al (2002), which was not detected in any collections from C. asparagi by comparison with an authentic standard. Complete identification and synthesis are in progress.

Figure 2: Mass spectrum of the candidate pheromone component from Crioceris asparagi

Key words: asparagus, Crioceris asparagi, pheromone, electroantennography, mass spectrometry

References Cossé, A., Bartelt, R, J. & Zilkowski, B. W. 2002: Identification and electrophysiological activity of a novel hydroxy ketone emitted by male cereal leaf beetles. J. Nat. Prod. 65: 1156-1160.

PheroFip 2019 155

SESSION 3

ALLELOCHEMICAL-BASED PEST

MANAGEMENT TACTICS

CHAIRPERSON: MICHELLE FOUNTAIN

PheroFip 2019 156

Investigating the role of leaf color and plant semiochemicals on the behaviour of Cacopsylla pyri

Bruna Czarnobai De Jorge1,3*, Hans E. Hummel2, Jürgen Gross1,3

1Julius Kühn-Institut, Federal Research Center for Cultivated Plants, Institute for Plant Protection in Fruit Crops and Viticulture, Laboratory of Applied Chemical Ecology, Schwabenheimer Str. 101, 69221 Dossenheim, Germany;2Justus-Liebig-Universität Gießen, Gießen, Germany; 3Technical University of Darmstadt, Chemical Plant Ecology, Schnittspahnstr. 10, Darmstadt, Germany *[email protected]

Abstract Worldwide are insects that can transmit diseases in the focus of research. In Europe, fruit trees of the family Rosaceae are seriously affected by phytoplasmas of the apple proliferation group. Phytoplasmas are plant pathogens causing a wide variety of symptoms that can cause serious symptoms and often lead to death of infected plants. Jumping plant lice of the genus Cacopsylla are their vectors. It is well known that they use chemical cues for orientation and host identification. To overcome the spreading of these diseases, especially the transition from infected to healthy plants, several techniques using volatile substances for attracting or repelling vector insects are being studied. In this study, we started to screen possible repellent substances like eugenol for repellence of Cacopsylla pyri, the pear leaf sucker. Secondly, we begun to evaluate the preferences of C. pyri for leaf colors from symptomatic and non- symptomatic pear plants, and to determine the relative effect of their spectral reflectance parameters for host finding.

Introduction Psyllids play a crucial role in the transmission of phytoplasmas from the AP group on fruit crops belonging to the Rosaceae (Tedeschi & Alma, 2004), as well as for transmission of Ca. Liberibacter (Lopes et al., 2009) on vegetables and Citrus plants. It was shown recently that these insects use chemical cues for orientation and host identification (Mayer et al., 2008a,b; Mann et al. 2012). The psyllid-Phytoplasma and the psyllid-Liberibacter systems have evolved convergently but share many similarities (Gross, 2016). Former studies indicate that Candidatus Phytoplasma pyri, a bacterium causing the pear decline disease, is able to overwinter in the body of C. pyri (Carraro et al. 2001; Garcia-Chapa et al. 2005), and, once infected, to spread the disease all over the vegetative period. Considering that the transmission of phytoplasma to plants occurs at the beginning of spring and that the symptoms of infection are more evident at the beginning of autumn, we suspect that on that period, the insects maybe more susceptible to acquire the bacteria from the plant. Therefore we want to determine the phytoplasma titer in C. pyri, for comparing its vector ability in samples collected in middle autumn and at the end of winter. As reported in recent years, the employment of attractive components like β- caryophyllene by creating insects traps lured for the capture of psyllids for monitoring purposes has been tested (Mayer et al. 2008a; Eben & Gross, 2013; Gross, 2014). As the infochemical produced by infected apple plants is attractive to both genders of vectoring psyllids, this could also enable the development of mass trapping systems for a more sustainable control of these insects in future. In the presented project, we want to investigate if (1) the leave color is influencing the behavior of Ca. pyri and (2) VOCs of infected or healthy trees could enable a mechanism of disease dispersion.

PheroFip 2019 157

Material and methods Olfactometer bioassays: Bioassays investigating the vector insect behavior to synthetic compounds have been carried out using dynamic Y-shaped olfactometer. By pumping purified air through two containers, one containing eugenol dissolved in methanol and the other containing just methanol as control, and attaching each outlet with a test arm of the olfactometer, the insect preference could be tested. The air flow was controlled with a flow meter. The tests were performed with females of the pear psyllid C. pyri. We were counting every individual that passed a final mark on one of the test arms within 5 min. To analyze data statistically dependent paired t–test was used. Spectral measurements: The optical properties of the leaves were acquired using a using a USB Ocean Optics FLAME-S-UV-VIS-ES and Xenon Pulse X2 lamp (Ocean Optics) light source. Each leaf was recorded from upside in the middle vein region. Reflectance properties were measured as the proportion of a diffuse reflectance standard (white standard). The fiber optics probe was mounted inside a matte black plastic tube to exclude ambient light and standardize the distance between each leaf probe at 1 cm. The angle of illumination and reflection was also fixed at 45° to minimize glare. Spectra were calculated at 5 nm intervals from 300 to 700 nm with SpectraSuite software. A black cardboard was used as the background of the leaf, thus enhancing the contrast features. Field trials: The field experiments were conducted in the experimental field at Julius Kühn- Institut, Dossenheim, Germany. All samples were stored directly after their collection in a freezer (-20 °C) for counting, sex and species determination. Beating tray samples were taken in October 2018 during the same period that we could identify symptoms of phytoplasma infection in plants on the field. Symptomatic trees are classified according to the color of the leaves which become abnormally red in autumn and start to drop prematurely. A white funnel cloth tray (50 × 50 cm) was held under a tree limb. Then, 10 limbs were beaten sharply three times each with a pole. Dislodged psyllids fell to the tray and were collected in plastic bags attached to the end of the funnel (Weintraub & Gross, 2013). Both symptomatic and healthy plants were sampled. Seventy-three trees were selected. Thirty-three were showing phytoplasma symptoms and forty not showing symptoms (five plants for row). We also investigated and counted both the eggs and nymphs (Pasqualini et al. 2003) on 10 leaves per tree. The samples were taken into the laboratory and examined using a binocular dissecting microscope.

Results and discussion Olfactometer bioassays showed that C. pyri females avoided the olfactometer arm containing eugenol. More than 72% of the tested females preferred the control arm (P < 0.01). Eugenol is a volatile found in a variety of plants including clove buds, cinnamon bark and leaves, tulsi leaves, turmeric, pepper, ginger, oregano and thyme. In addition, several other aromatic herbs including basil, bay, marjoram, mace and nutmeg are also claimed to have significant quantity of eugenol (Raja et al. 2015). Clove essential oil has been widely studied for its insecticidal and repellent activities against many species of pests (Chaieb et al. 2007, Kafle & Shih, 2013, Corte´s-Rojas et al. 2014), such as Asian citrus psyllid (Mann et al., 2010). As reported by Tian et al. (2015), clove essential oil treatment reduced the numbers of C. chinensis nymphs in a concentration-dependent manner. These results provided us indication that the major components of clove essential oil (eugenol) could be repellent and/or deterrent on female psyllids searching for an appropriate oviposition site. More studies will be carried to investigate the potential of those volatiles as behavior modifying. The field of insects repellents is moving forward as farmers and consumers demand means of protection from arthropod that are safe to use and environmentally sustainable. Perhaps the most important

PheroFip 2019 158

consideration is improving the longevity of those repellents that are effective but volatile. Hence, we are conducting studies to improve formulations and as well increase volatiles longevity. The spectral measurements showed that not symptomatic leaves had higher reflectance mean in the range of 482 -585 nm (blue, green and yellow, P < 0.01). On the other hand, symptomatic leaves had higher reflectance in the range of 585 - 700 nm (orange and red, P < 0.01). Field samples of adults showed that the only species found on the field was C. pyri, and the majority was winterforms (Garcia-Chapa et al. 2005). The mean number of eggs laid on symptomatic trees was ten times lower than on infected ones (P < 0.01). The numbers of nymphs showed no differences on symptomatic and non-symptomatic trees. In the same period, we could not find differences between the mean number of adults collected on plants with and without phytoplasma symptoms. The importance of visual stimuli in the behavior of psyllids and its response to light and trap color has been reported for D. citri (Hall et al. 2007, Wenninger et al. 2009, Sétamou et al., 2012). Hall et al. (2010) have evaluated the preferences of different hues of green and yellow traps. At the peak period of D. citri flight, the red and the yellow traps had comparable D. citri captures in the lemon grove. The blue and white traps captured the fewest numbers of D. citri adults throughout the study (Sétamou et al. 2014). Nevertheless, the importance of visual signals for pear psyllids remains unclear. Combining the results of spectral measurements with data from VOCs collection of infected trees will enable us to assess the role of volatiles and leaf color in the pear psyllids/pear decline disease system. The presented results are part of a pioneer project, which consists the development of new types of repellent formulations and different approaches such as attractive colors for developing efficient traps. In that field, nanotechnology can be applied for agrochemicals by using nanoscale carriers; they have controlled release mechanisms which allow the active ingredient to be taken up slowly, thus improving its effectiveness, while reducing the amount applied. Against the background of insect behavior in the field, we want to establish new techniques of vector control for prevention of new phytoplasma infections.

Key words: apple proliferation phytoplasma, push-and-pull, psyllids, application, infochemicals, chemical ecology, kairomones, allomones, pheromones, reflectance, repellents

Acknowledgements We thank CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico, Brasil) for financial support.

References Chaieb, K., H., Hajlaoui, T., Zmantar, A. B., Kahla-Nakbi, M., Rouabhia, K., Mahdouani, & A. Bakhrouf. 2007: The chemical composition and biological activity of clove essentialoil, Eugenia caryophyllata (Syzigium aromaticum L. Myrtaceae): a short review. Phytother. Res. 21: 501–506. Cortés-Rojas, D. F., de Souza, C. R. F., & Oliveira, W. P. 2014: Clove (Syzygium aromaticum): a precious spice. Asian Pacific journal of tropical biomedicine 4(2): 90-96. Eben, A., & Gross, J. 2013: Innovative control of psyllid vectors of European fruit tree phytoplasmas. Phytopathogenic Mollicutes 3(1): 37-39. Garcia-Chapa, M., Sabaté, J., Laviña, A., & Batlle, A. 2005: Role of Cacopsylla pyri in the epidemiology of pear decline in Spain. European Journal of Plant Pathology 111(1): 9- 17.

PheroFip 2019 159

Gross, J., 2011: Chemical ecology of psyllids and their interactions with vectored phytoplasma and plants. IOBC WPRS Bulletin 72: 123-126. Gross, J., 2014: Research on Chemically Mediated Communication between Cultivated Plants and Pest Organisms–Basis for Innovative Applications in Phytomedicine (Doctoral dissertation). Gross, J., 2016: Chemical communication between phytopathogens, their host plants and vector insects and eavesdropping by natural enemies. Front. Ecol. Evol. 4: 104. doi: 10.3389/fevo.2016.00104. Hall, D. G., Sétamou, M., & Mizell III, R. F. 2010: A comparison of sticky traps for monitoring Asian citrus psyllid (Diaphorina citri Kuwayama). Crop Protection 29(11): 1341-1346. Hall, D. G., Hentz, M. G., & Ciomperlik, M. A. 2007: A comparison of traps and stem tap sampling for monitoring adult Asian citrus psyllid (Hemiptera: Psyllidae) in citrus. Florida Entomologist 327-334. Horton, D. R. 1994: Relationship among sampling methods in density estimates of pear psylla (Homoptera: Psyllidae): implications of sex, reproductive maturity, and sampling location. Ecology and Population Biology 87: 583-591. Kafle, L. & C. J. Shih. 2013: Toxicity and repellency of compounds from clove (Syzygium aromaticum) to red imported fire ants Solenopsis invicta (Hymenoptera: Formicidae). J. Econ. Entomol. 106: 131–135 Lopes, S. A., Bertolini, E., Frare, G. F., Martins, E. C., Wulff, N. A., Teixeira, D. C., Fernandes, N. G. & Cambra, M. 2009: Graft transmission efficiencies and multiplication of “Candidatus Liberibacter americanus” and “Ca. Liberibacter asiaticus” in citrus plants. Phytopathology 99: 301–306. doi: 10.1094/PHYTO-99-3-0301. Maia, M. F. & Moore, S. J. 2011: Plant-based insect repellents: a review of their efficacy, development and testing. Malaria journal 10(1): S11. Mann, R. S., Ali, J. G., Hermann, S. L., Tiwari, S., Pelz-Stelinski, K. S., Alborn, H. T. & Stelinski, L. L. 2012: Induced release of a plant-defense volatile ‘deceptively’attracts insect vectors to plants infected with a bacterial pathogen. PLoS pathogens 8(3): e1002610. Mayer, C. J., Vilcinskas, A. & Gross, J. 2008a: Pathogen-induced release of plant allomone manipulates vector insect behaviour. Journal of Chemical Ecology 34: 1518- 1522. Mayer, C. J, Vilcinskas, A. & Gross, J. 2008b: Phytopathogen lures its insect vector by altering host plant odor. J Chem Ecol 34:1045–1049 Pasqualini, E., Civolani, S. & Corelli Grappadelli, L. 2003: Particle film technology: approach for a biorational control of Cacopsylla pyri (Rhynchota: Psyllidae) in Northern Italy. Bulletin of Insectology 55: 39-42. Raja, M. R. C., Srinivasan, V., Selvaraj, S., & Mahapatra, S. K., 2015: Versatile and synergistic potential of eugenol: a review. Pharm Anal Acta 6(5): 367. Sétamou, M., Sanchez, A., Patt, J. M., Nelson, S. D., Jifon, J., & Louzada, E. S. 2012: Diurnal patterns of flight activity and effects of light on host finding behavior of the Asian citrus psyllid. Journal of Insect Behavior 25(3): 264-276. Sétamou, M., Sanchez, A., Saldaña, R. R., Patt, J. M., & Summy, R. 2014: Visual responses of adult Asian citrus psyllid (Hemiptera: Liviidae) to colored sticky traps on citrus trees. Journal of insect behavior 27(4): 540-553. Tedeschi, R., & Alma, A. 2004: Transmission of apple proliferation phytoplasma by Cacopsylla melanoneura (Homoptera: Psyllidae). Journal of Economic Entomology 97(1): 8-13

PheroFip 2019 160

Tian, B. L., Liu, Q. Z., Liu, Z. L., Li, P. & Wang, J. W. 2015: Insecticidal potential of clove essential oil and its constituents on Cacopsylla chinensis (Hemiptera: Psyllidae) in laboratory and field. Journal of economic entomology 108(3): 957-961. Weintraub, P. G. & Gross, J. 2013: Capturing insect vectors of phytoplasmas. In Dickinson M.J. & Hodgetts, J. (eds.) Phytoplasma: Methods and Protocols. Methods in Molecular Biology 938: 61-72. Springer Science + Business Media, New York. Wenninger, E. J., Stelinski, L. L. & Hall, D. G. 2009: Role of olfactory cues, visual cues, and mating status in orientation of Diaphorina citri Kuwayama (Hemiptera: Psyllidae) to four different host plants. Environ Entomol 38: 225–234.

PheroFip 2019 161

KLIMAKOM – Insect's communication under climate change

Margit Rid1*, Christine Becker2, Annette Reineke2**, Jürgen Gross1,3***

1Julius Kühn-Institut, Federal Research Centre for Cultivated Plants, Institute for Plant Protection in Fruit Crops and Viticulture, Laboratory of Applied Chemical Ecology, Schwabenheimer Str. 101, Dossenheim, Germany; 2Geisenheim University, Department of Crop Protection, Von-Lade-Str. 1, Geisenheim, Germany; 3Technical University of Darmstadt, Chemical Plant Ecology, Schnittspahnstr. 10, Darmstadt, Germany *[email protected]; **[email protected]; ***[email protected]

Abstract Climate change can threaten our food production. Because of changing temperatures and concentrations of greenhouse gases, well-established pest insect control techniques may have to be adapted to the changing requirements to stay effective. Especially temperature, elevated carbon dioxide (CO2) and ozone concentrations will have significant impacts on the environment (IPCC, 2014) and therefore certainly on control strategies based on chemical ecology of insects. The aims of the project are the investigation of effects of these fundamental determinants of climate change on the perception and reaction of insects to semiochemicals, as well as on the stability and persistence of volatile organic compounds (VOCs) in the environment. This fundamental research will create the basis for the identification of possible limitations for plant protection methods based on semiochemicals, and to adapt them to a changing environment. Behavior modifying compounds (pheromones, attractants and repellents) will be evaluated in lab, greenhouse and field experiments under different environmental conditions (temperature, elevated CO2, and O3 concentrations) regarding their effects and efficiency on several insect pest species. The emphasis is set on the mating disruption method which is widely used in Europe for the biological control of our first model organism Lobesia botrana (European grapevine moth) (Fig. 1). This includes investigation of the stability of the main component of its sexual pheromone ((7E,9Z)-dodecadienyl acetate (E7,Z9-12Ac)) (Roelows et al. 1973), as well as the release and reception of E7,Z9-12Ac and minor pheromone components by conspecifics. Chemical analysis including gas chromatography coupled with mass spectrometry, electrophysiological methods as GC-EAD, behavioral tests in wind tunnel, olfactometer and field cages will be used. The second model organism investigated in this project are jumping plant lice of the genus Cacopsylla (Heteroptera: Psyllidae), which are well known as vectors of phytoplasma diseases of fruit crops (Jarausch et al. 2018). Psyllids are phloem feeders and both nymphs and adults feed on plant sap. Phytoplasma vector species on apple and stone fruits are univoltine whereas pear psyllids are mostly polyvoltine with overlapping generations (Jarausch et al. 2018). Polyvoltine vector species usually overwinter as adults on or near to their host plants, while univoltine species have an obligate host alternation and migrate in summer (= emigrants) to their overwintering plants (conifers), returning to their respective reproduction host plants in spring (= remigrants) (Mayer et al. 2011; Burckhardt, 1994).

PheroFip 2019 162

Figure 1: Stages of insect pheromone communication and factors that could affect them.

Their migration behavior, their search for appropriate food and oviposition sites are regulated by plant VOCs (Gross, 2016) and non-volatile phloem constituents (Gallinger & Gross, 2018), which differ between plant species and may be influenced by psyllid feeding, phytoplasma infection as well as further biotic and abiotic factors (Fig. 2). The influence of the fundamental determinants of climate change on these multitrophic interactions will be the second emphasis of the study. The main goal of the project is suitable for the improvement of plant protection measures. We will focus on two key aspects pursuing the following goals: 1. Reduction of insecticide spraying in fruit crops and viticulture through non-chemical plant protection measures adapted to desideratum of climate change (mating disruption, monitoring, mass trapping). 2. Reduction of new infections of fruit trees with phytoplasma diseases by improved monitoring and selective and targeted control of their vectors. Here we introduce our new project in which we investigate if greenhouse gases and reactive oxygen species have the potential to alter semiochemically mediated communication within and between insects and their host plants and transmitted pathogens. Our hypothesis is that both pheromone and allelochemical-mediated communication is influenced by the determinants of climate change. We address three main questions: Does the chemical structure of the semiochemicals is changed by climate change determinants and/or are both the emitters (insects, plants) or the receivers influenced. We invite all researchers attending the congress to discuss our experimental setting and preliminary results with us. At this point of our project we appreciate as much as input as possible.

PheroFip 2019 163

Figure 2: Stages of psyllid interspecific communication during migration between alternate hosts and factors that could affect them.

Key words: ozone, carbon dioxide, elevated temperature, chemical communication

Acknowledgement The project is supported by funds of the Federal Ministry of Food and Agriculture (BMEL) based on a decision of the Parliament of the Federal Republic of Germany via the Federal Office for Agriculture and Food (BLE) under the innovation support program.

References Burckhardt, D. 1994: Psylloid pests of temperate and subtropical crop and ornamental plants (Hemiptera, Psylloidea): a review. Entomology (Trends in Agricultural Science) 2: 173– 186. Gallinger, J. & Gross, J. 2018: Unraveling the host plant alternation of Cacopsylla pruni – Adults but not nymphs can survive on conifers due to phloem/xylem composition. Front Plant Sci 9: 484. doi: 10.3389/fpls.2018.00484. Gross, J. 2016: Chemical communication between phytopathogens, their host plants and vector insects and eavesdropping by natural enemies. Front. Ecol. Evol. 4: 104. doi: 10.3389/fevo.2016.00104.

PheroFip 2019 164

IPCC. 2014: Climate Change 2014: impacts, adaptation, and vulnerability. Part A: global and sectoral aspects. Contributions of working group II to the fifth assessment report of the intergovernmental panel on climate change. Cambridge University Press, Cambridge. Jarausch, B., Tedeschi, R., Sauvion, N., Gross, J. & Jarausch, W. 2018: Psyllid vectors. In: Phytoplasmas: plant pathogenic bacteria, Volume 2: “Transmission and management of phytoplasma associated diseases”. (Editors: A. Bertaccini, P. Weintraub, G.P. Rao, N. Mori). In press Mayer, C. J., Vilcinskas, A. & Gross, J. 2011: Chemically mediated multitrophic interactions in a plant-insect vector-phytoplasma system compared with a partially nonvector species. Agric For Entomol 13: 25–35. Roelofs, W. L., Kochansky, J., Cardé, R. T., Arn, H. & Rauscher, S. 1973: Sex attractant of the grapevine moth, Lobesia botrana Mitt. Schweiz. Entomol. Ges. 46: 71-73.

PheroFip 2019 165

Luring aphid hyperparasitoids in sweet pepper greenhouses

Jetske G. de Boer1, Louise E.M. Vet1,2

1 Netherlands Institute of Ecology (NIOO-KNAW), Department of Terrestrial Ecology, PO Box 50, 6700, AB, Wageningen, The Netherlands; 2Laboratory of Entomology, Wageningen University, PO Box 16, 6700 AA, Wageningen, The Netherlands

Abstract Aphids are one of the main pests in protected cultivation of sweet pepper. A range of natural enemies is available as biological control agents, including braconid parasitoids of the genus Aphidius. However, Aphidius also has natural enemies in the greenhouse environment, which can reduce the efficiency of biological control. Mummified aphids containing the (pre)pupae of Aphidius parasitoids are often attacked by hyperparasitoids of the familie Megaspilidae and Pteromalidae. Offspring of these hyperparasitoids develop inside the mummy and kill the primary parasitoid, thus preventing establishment of parasitoid populations and indirectly favouring aphid populations. In organic sweet pepper greenhouses in The Netherlands, hyperparasitism rates can go up to 100%, depending on the time of the year. In this project, we aim to control hyperparasitoids of aphid parasitoids in sweet pepper crops by developing lures (baits) that divert hyperparasitoids away from their beneficial parasitoid host. To develop these lures, we study host-searching behaviour of hyperparasitoids, with a focus on Dendrocerus aphidum. This species appears to be the most common hyperparasitoid in sweet pepper greenhouses. Behavioural assays (2-chamber olfactometer and Y-tube olfactometer) have shown that mummies are attractive to D. aphidum and congeneric species. Plant volatiles may also play a role in host searching behaviour over longer distances. These findings show the potential of using attractive odours to trap aphid hyperparasitoids in greenhouses, and headspace entrainment and chemical analyses of the attractive odour sources are currently done.

Key words: hyperparasitoid wasp, volatiles, behavioural ecology, chemical ecology, aphid biocontrol

PheroFip 2019 166

Is globulol mediating attack by male pioneers of Gonipterus platensis (Col., Curculionidae)?

Sofia Branco1, Eduardo Mateus1, Stefan Schütz2, Maria Rosa Paiva1

1CENSE- Center for Environmental and Sustainability Research, DCEA, Faculdade de Ciências e Tecnologia (FCT),Universidade Nova de Lisboa (UNL),Caparica Portugal; 2Department of Forest Zoology and Forest Conservation, Buesgen-Institute Göttingen University Göttingen Germany

Abstracts Weevils of the genus Gonipterus (Col., Curculionidae feed on eucalyptus young leaves, buds and shoots, causing reduced tree growth and mortality. Gonipterus platensis is the main coleopteran defoliator of eucalyptus worldwide, being also present in Spain and Portugal, where economic damage occurs particularly at high altitudes, where above 700m mean defoliation values can attain 75% (Reis et al. 2012). The process of tree selection and colonization by G. platensis has not yet been clarified. Although Gonipterus spp. feed exclusively on eucalyptus foliage, preferences for one particular species over another have been commonly reported. Furthermore, the weevils also preferentially select different clones of the same eucalyptus species, even when planted on the same sites (e.g. Paiva & Mateus, 2012). In an Eucalyptus globulus stand, located near Lisbon, Loures municipality (38°49'19"N, 9°12'23"W), a preliminary study was conducted to compare the volatiles emitted by the foliage of trees that were subjected to either mechanical damage, to damaged by herbivory, or were left undamaged. Before sampling, trees were thoroughly inspected for signs of the characteristic damage caused by G. platensis. Three undamaged trees and three trees showing signs of herbivory – chewed leaves, were selected and one branch was collected from each tree. Three treatments were applied, namely: i) Undamaged leaves, from undamaged branches; ii) Chopped (mechanically damaged) leaves, from undamaged branches; iii) Chewed leaves. For each treatment, three samples consisting of two newly formed leaves were prepared and the volatiles emitted were collected by head space micro-extraction and analysed by gas chromatography mass spectrometry. Results indicated that some compounds were only detected in damaged leaves and that, in parallel, emissions of most VOCs increased with damage. Surprisingly, globulol was emitted in much lower amounts by herbivory damaged leaves, than by undamaged and mechanically damaged leaves – Fig. 1. Additionally, previous studies using GC-MS-EAD showed that globulol was detected by the antennae of both male and female weevils - Fig. 2. Furthermore, in olfactometer trials, globulol triggered a repellent effect in virgin males. Yet, no response was elicited from either mated males or females (Branco et al. 2018). Since globulol is a common compound present in the emissions of eucalyptus species, two alternative hypothesis might be considered to explain findings: i) G. platensis could preferentially select trees with lower globulol levels, possibly due to a reduced capacity to detoxify this compound, known to have antimicrobial properties; ii) Alternatively, inoculation of saliva by the weevils while feeding, might interrupt the cascade mechanism of globulol production. This could signal weevils that a certain tree is already under attack by conspecifics. Further research is required.

PheroFip 2019 167

1.6E+08

1.4E+08

1.2E+08

1.0E+08

8.0E+07

6.0E+07

Ionarea 109 4.0E+07

2.0E+07 0.0E+00 herbivory undamaged mechanical damage damage

Sampled leaves (average)

Figure 1: Emission of globulol by E. globulus leaves. Bars represent the mean integrated chromatographic areas of ion 109 obtained for each SPME treatment: herbivory damaged, undamaged and mechanical damaged.

2e+6 515

510 2e+6

505

1e+6

EAG mV 500

GC peak intensity (TIC) 5e+5 495

0 490 19 20 21 22 23 24 25 Retention Time (min)

Figure 2: GC-MS chromatographic profile of E. globulus (bottom trace) on an INNOWAX column and corresponding EAG recording (top trace) obtained with a female antenna, the peak corresponding to globulol is marked with an *

Key words: Eucalyptus globulus, Gonipterus platensis, host selection, globulol

Acknowledgements This work was partially supported by CENSE (Center for Environmental and Sustainability Research) which is financed by national funds from FCT/MEC (UID/AMB/04085/2013).

PheroFip 2019 168

References Branco, S., Mateus, E. P., da Silva, M. D. G., Mendes, D., Rocha, S., Mendel, Z., Schütz, S. & Paiva, M. R. 2018: Electrophysiological and behavioural responses of the Eucalyptus weevil, Gonipterus platensis, to host plant volatiles. Journal of Pest Science 1-15. Paiva, M. R. & Mateus, E., 2012: Protecção do eucalipto contra o gorgulho do eucalipto “Gonipterus I” - Relatório final, Lisboa. Reis, A. R., Ferreira, L., Tomé, M., Araujo, C. & Branco, M. 2012: Efficiency of biological control of Gonipterus platensis (Coleoptera: Curculionidae) by Anaphes nitens (Hymenoptera: Mymaridae) in cold areas of the Iberian Peninsula: implications for defoliation and wood production in Eucalyptus globulus. Forest Ecology and management 270: 216-222.

PheroFip 2019 169

Attract and Kill for the control of olive fruit fly in Alto Garda Trentino

Massimo Mucci1*, Mario Baldessari1, Franco Michelotti1, Serena Giorgia Chiesa1, Gino Angeli1

1Technology Transfer Center, Fondazione Edmund Mach, V. E. Mach, 1 38010 S. Michele a/A, Italy *[email protected]

Abstract Alto Garda Trentino, in northern Italy, is an important economic touristic area characterized by the insubric climate typical of Lake Garda basin. Good quality oil production came from the local olive groves and the olive fruit fly (Bactrocera oleae, Rossi) represent the most serious and potentially devastating treat. IPM as control strategy was largely adopted with positive results but during the last decade the frequency of the B. oleae heavy attacks increased generating a general concern. The technological-scientific progress, the social environmental issues awareness, the massive use of chemical pesticides and their regulations enforced by the laws were drivers of the pest management. One objective of the project named “Innovazione e Ricerca per l'Olio Extravergine dell'Alto Garda Trentino” funded by Agraria Riva del Garda and Provincia Autonoma di Trento, was the transfer of knowledge about the reliability strategies for the management of the olive fruit fly with modern tools. A three years experiment (2016-2018) was carried out to evaluate the effectiveness of different Attract and Kill approaches, mass trapping and “Lure and Kill" tactics. During 2016 and 2017 were compared two “Lure and Kill” bait station products; Eco-Trap (Vioryl), a bag treated with contact insecticide (deltamethrin) containing ammonium bicarbonate and provided with a sexual pheromone (1,7-dioxaspiro[5.5]undecane) dispenser and Spintor Fly (Dow AgroSciences), a sprayable bait protein-specific with ingestion insecticide (spinosad). In 2017 it was added to compare a trap for mass trapping made of a conic plastic body containing specific food and sex attractant and a cover internally treated with deltamethrin, Flypack (SEDQ). The insects inside traps were counted to follow density of the population. In 2018 Flypack and Eco-Trap comparison was repeated. The trial was set up in a hilly olive Casaliva orchard (around 100 hectares) located in monte Brione (Riva del Garda, TN). The experimental design, according to European Plant Protection Organisation (EPPO) standards, consisted of adjacent large plots per types of treatment (around 3 hectares/plot) subdivided in three subplots size (at least 1 hectare each with a net areas of 0.1 hectare or 20 trees at least, separated by a minimum of 100 m) at different altitude. Untreated plots were selected outside the trial area with conditions closely similar. The experiments started every year in July during the phenological phase of pit hardening (BBCH 75), when the olives were susceptible to egg laying, until harvest. The Eco-Trap and Flypack were hung with a density of 100 and 70 traps per hectare/year respectively. Spintor Fly was applied before the fruit first punctures and following the insect infestations (with maximum of eight treatments/year) at dosage of 1 L of product diluted in 4 L water/hectare (half of total number of the trees were treated on a small part of foliage by backpack sprayer). The first year of study, 2016, was characterized by average of 30% of olives total infested on the untreated area. Spintor Fly, in particular, with eight applications during season, showed the best protection results (6% fruit damaged). In the next two years the damages at harvest in the untreated trial area gradually lowered (from 24% to 15% in 2017 and 2018 respectively) and all products tested (Eco-Trap, Spintor Fly and Flypack) showed similar good results (below of 5% olives total infested).

PheroFip 2019 170

Attract and Kill, as alternative IPM, solved in part important problems about pollution, residues, resistance, drift and side effects on beneficial. From our experience some considerations can be made; the best results were obtained in case of low infestations and when the applications were repeated during years. It needed area-wide applications to guarantee success of the technique as replacement of insecticides based strategy. Spintor Fly provided good protection, use flexibility and fewer residues on the fruit but it was necessary consider its cost, risk of wash off and low persistence. The Eco-Trap allowed adequate protection and when it needed, especially during seasons with high infestations and near harvest when olives was more vulnerable to attack, could be combined with bait spray (e.g. Spintor Fly) or insecticide applications. Flypack, when commercially available, for mass trapping could represent a valid alternative to chemical insecticides and bait applications especially in heavy anthropized and touristic sites where treatments were not possible. When the risk of immigration of mated females was low as in our study area, we could confirm Flypack to be a useful device for spring captures of overwintering adult flies lowering their potential of generating the future population. A constant B. oleae monitoring and the adoption of the good agronomic practices, such as anticipating the harvest in case of high infestations, still remain of fundamental importance for preservation of the good effectiveness results obtained with the Attract and Kill method.

Key words: olive fruit fly, Integrated Pest Management, semiochemical-based control

PheroFip 2019 171

Morphology and ultrastructure of the antennal sensilla of Sitophilus granarius (COLEOPTERA: CURCULIONIDAE)

Suliman A. I. Ali1,2*, Mory Mandiana Diakite1, Saqib Ali, Man-Qun Wang1*

1Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, People’s Republic of China; 2Agricultural Research Corporation (ARC), P. O. Box 126, Wad Medani, Sudan. Entomology Research Section *[email protected]

Abstract Sensilla are sense organs in insects, typically consisting of a group of cuticle or epidermal cells that appear as hairs or rod-shaped structures. Sensilla serve as the functional elements of sensory systems. The goal of this study was to determine the type and distribution of sensilla in the antennae of Sitophilus granarius (L.) using light microscopy, and scanning and transmission electron microscopy (TEM). This is one of the first studies in which the morphology and distribution patterns of sensilla on antennal segments of male and female S. granarius have been investigated using scanning electron microscopy, followed by TEM. Different morphological sensilla types, including two sensilla basiconica types, with two subtypes, four sensilla coeloconica types and five sensilla chaetica types, with one subtype, have been identified on S. granarius antennae, whose external structure and shape are peculiar. TEM micrographs of sensilla basiconica on the antennae of S. granarius are characterized by strongly corrugated pores around the cuticle, while micrographs of sensilla coeloconica longitudinal sections showed flat-tipped and smooth-surfaced pegs bearing an apical pore that is suggestive of a gustatory function. TEM micrographs of sensilla chaetica longitudinal sections showed dendrite branches and cuticular pore arrow heads that may be involved in the perception of humidity, temperature, heat and CO2. Because sensilla chaetica of different sizes were distributed around the head and rostrum, these may function as contact- chemoreceptors .These results are discussed in relation to the possible roles of the sensilla types in the host location behavior of S. granarius

Key words: Sitophilus granarius; antenna; sensilla; scanning electron microscopy; transmission electron microscopy

PheroFip 2019 172

SESSION 4

PHEROMONE-BASED PEST

MANAGEMENT TACTICS

CHAIRPERSON: ALLY HARARI

PheroFip 2019 173

Measurement of Lobesia botrana airborne pheromone by high volume air sampling in mating disruption-treated vineyards

Aitor Gavara1, Ismael Navarro2, Sandra Vacas1, Jaime Primo1, Vicente Navarro-Llopis1

1CEQA-Instituto Agroforestal del Mediterráneo, Universitat Politècnica de València, Camino de Vera s/n, edificio 6C-5ª planta, 46022 Valencia (Valencia), Spain; 2Ecología y Protección Agrícola SL, Pol. Ind. Ciutat de Carlet, 46240 Carlet (Valencia), Spain

Abstract Lobesia botrana (Denis & Schiffermüller) (Lepidoptera: Tortricidae) is a key pest in vineyards of southern Europe. Pheromone-based mating disruption against this pest is widely used due to their effectivity and environmental benefits compared with the use of pesticides. Pheromone cost is a deciding factor for the implementation of this technique; for this reason, knowledge of the airborne pheromone quantity required to trigger the disruption effect is essential. Developing precise and reproducible air sampling and pheromone quantification protocols will help us to adjust the exact required quantity of pheromone to produce the disruption effect. For this purpose, we used a high volume air sampler to collect air samples in field at three different moments of the crop season (beginning, second pest peak and pre- harvest) to detect and measure airborne pheromone concentrations. Pheromone was captured in polyurethane foam-resin adsorbent sandwiches and the resulting solvent extracts were analyzed by gas chromatography coupled to triple quadrupole mass spectrometry for pheromone quantification. These measures were performed in vineyards treated with two different disruption methods: passive pheromone dispensers and aerosol devices. These data will allow us to optimize the application of the mating disruption technique in order to reduce total costs and to understand the disruption mechanisms.

Key words: European grapevine moth; Lepidoptera; mating disruption; sex pheromone

PheroFip 2019 174

Geostatistical Approach for Spatial Distribution Analysis of Lobesia botrana (Den. & Schiff): (Lepidoptera: Tortricidae) in Douro Demarcated Region (DDR)

J. Salvação1, C. Carlos2;3, A. Ferreira3, M. Nóbrega4, G. Fonseca4, J. Oliveira5, D. Gomes6, S. Soares7, A. Martinho7, R. Soares7, F. Gonçalves2, L. Torres2, J. Aranha2

1UTAD/ECAV - Scholarship Researcher of Operational Group - CSinDouro, University of Trás-os- Montes and Alto Douro - School of Agrarian and Veterinary Sciences, 5001-801 - Vila Real, Portugal 2CITAB – Centre for the Research and Technology of Agro-Environmental and Biological Sciences, University of Trás-os-Montes and Alto Douro, 5001-801 - Vila Real, Portugal; 3 ADVID – Association for the Development of Viticulture in the Douro Region, Quinta de Santa Maria, Apt. 137, 5050-106 - Godim, Portugal; 4SOGEVINUS FINE WINES S.A., Av. Diogo Leite, 344 - 4400-111 - Vila Nova de Gaia, Portugal; 5DONA MATILDE VINHOS LDA., R. Nova da Alfândega, 7, sala 204 4050-430 - Porto, Portugal; 6QUINTA DO VALLADO SOCIEDADE AGRÍCOLA LDA., Vilarinho dos Freires, 5050-364 - Peso da Régua, Portugal REAL COMPANHIA VELHA, R. Azevedo Magalhães 314, 4430- 022, Vila Nova de Gaia, Portugal

Abstract The European grapevine moth (EGVM), Lobesia botrana (Den & Schiff.) is the most important vineyard pest in Douro Demarcated Region (DDR) (North of Portugal) where the pest typically develops three generations per year, of which the third is the most damaging one. The mating disruption (MD) technique, an environmental safe control method against this pest was first used in this region in 2000, being increasingly applied since then. However, within this steep slope viticulture region, some major constraints to use of MD techniques were previously identified. The DDR is characterised by a very fragmented landscape with the majority of the vineyards organized in small plots, often bounded by other crops such as olive groves and by unmanaged non-crop habitats. The climatic conditions in DDR allow a high biotic potential for EGMV development. The strong altitude range and rugged orography as well large number of plots under MD treated are important issue to consider for the successful application of the technique, in order to ensure the homogeneous distribution of the pheromone cloud through the treated area. To overcome some of these constraints, since the end of 2017 a partnership was set up between ADVID, a winegrower organization, the University of Trás-os-Montes e Alto Douro (UTAD) and four wine companies (Real Companhia Velha, Sogevinus Fine Wines, S.A., Dona Matilde Vinhos Lda. e Quinta do Vallado Sociedade Agrícola Lda.) through the project “CSinDouro - Confusão sexual (CS) contra a traça-da-uva, Lobesia botrana (Denn. & Schiff.) em viticultura de montanha: caso particular da Região Demarcada do Douro (RDD)”. A key objective of this project is to investigate L. botrana population spatio-temporal dynamics inside and outside vineyards plots, in order to evaluate the effect of the landscape elements on pest distribution and the effectiveness of MD. To introduce improvements in the use of MD at the plot level, in DDR conditions, a characterization of spatio-temporal variability, on adult male catches in pheromone traps, was performed through Geostatistical methods (IDW - Inverse Distance Weighting and Kriging), as well as an estimation of EGVM damages, by plot and by farm, in the three pest generation. The achieved results will be used for improving pheromone dispenser location as a way to get a homogeneous distribution of the pheromone cloud, according to landscape characteristics.

Key words: Douro Demarcated Region, Geostatistical Analysis, Lobesia botrana, Mating Disruption, Spatial Distribution, Vineyard

PheroFip 2019 175

Development of the dual purpose mating disruption dispenser to control both the European grapevine moth, Lobesia botrana, and the vine mealybug, Planococcus ficus, in vineyards

Akihiro Baba1, Erina Ohno1, Tatsuya Hojo1, Ryuichi Saguchi1, Takeshi KIinsho1

1Specialty Chemicals Research Center, Shin-Etsu Chemical Co., Ltd., Niigata, Japan

Abstract The European grapevine moth (EGVM), Lobesia botrana (Lepidoptera: Tortricidae), and the vine mealybug (VMB), Planococcus ficus (Hemiptera: Pseudococcidae), are serious pests in vineyards. From the point of view of IPM, pheromone-based mating disruption (MD) is one of the most effective methods for controlling these pests. Regarding the control of EGVM, greater success has been achieved in areas where Shin-Etsu dispenser ISONET-L is applied. ISONET-PF for controlling VMB has also proved the good efficacy in MD trials, and is now ready to launch. ISONET-L and ISONET-PF are promising tools in order to control EGVM and VMB, and can be used alone or in combination depending on the pest presence. To control both EGVM and VMB more conveniently and eco-friendly, the dual purpose MD dispenser, ISONET-LPF has been developed. Many factors should be considered for the development of ISONET-LPF, including: active ingredient (AI) dose, dispenser raw material, AI stability, release rate, longevity, stock stability, weather resistance in field, industrial manufacturability and cost. Field MD trials using ISONET-LPF have been conducted in Italy and Spain. MD efficacy was demonstrated through suppression of the male trap capture and reduction of the damage.

Key words: European grapevine moth, Lobesia botrana, vine mealybug, Planococcus ficus, mating

PheroFip 2019 176

Control of oriental fruit moth, Cydia molesta Busck and peach twig borer Anarsia lineatella Zell. using reduced rate of pheromone dispensers

Hristina Kutinkova1, Vasiliy Dzhuvinov1, Desislava Stefanova1, Radoslav Andreev2, Nedyalka Palagacheva2, Bill Lingren3

1Fruit Growing Institute, 12 Ostromila str., 4004 Plovdiv, Bulgaria ([email protected]) 2Agricultural University, “Mendeleev” bul.12, 4000 Plovdiv, Bulgaria 3Trécé Inc., Adair OK, USA

Abstract Peach is one of the major fruit species in Bulgaria. It provides rapid return of investments due to early bearing and relatively low phytosanitary problems. Its main pest is the oriental fruit moth (OFM), Cydia molesta (Busck) (Lepidoptera: Tortricidae). The larvae of early OFM generations damage current season shoot tips; then they feed in the developing fruitlets and fruits. Peach twig borer (PTB) Anarsia lineatella (Zell.) (Lepidoptera: Gelechiidae) is an Eurasian species that has spread extensively worldwide; its larvae can develop on many plant species from the genera Cydonia, Malus, Prunus and Pyrus. In Bulgaria, the main hosts are Prunus armeniaca L. (apricot) and Prunus persica L. (peach). Chemical pest control in peach and nectarine fruit orchards of Bulgaria has consistented of a broad spectrum of organophosphate and pyrethroid insecticides. Recently their effectiveness has decreased, apparently due to the development of resistance in the pest. However, this is not well documented. Although quite effective, environmental problems and consumer concerns have arisen. Deregistration of many of the insecticides used in the control of these insects along with public demand for residue free products, has increased the interest for innovative tools in pest management. Behavior modifying pheromones can be used for environmentally safe insect management. And the technique has become one of the most important treatments for controlling the main insects in orchards. Mating disruption technology has been successfully used for control of oriental fruit moth and peach twig borer. The trials were carried out in the years 2016 - 2018 in an isolated peach experimental orchard of 1 ha in the Fruit Growing Institute, Plovdiv – Central South Bulgaria. Mating disruption (MD) was tested as an alternative method for controlling oriental fruit moth, Cydia molesta (OFM) and Anarsia lineatella (PTB) from post-bloom till harvest. CIDETRAK® OFM/PTB MESO™ pheromone dispensers of Trécé Inc., USA were installed once during the season and were hung in the upper third of the tree canopy at a density of 20 dispensers per ha, before the start of OFM and PTB flights. These dispensers are designed to deliver a long-lasting performance for the whole season, with remarkably fast application. Against other pests occurring in the trial plots, aphicide treatments (two per season) were applied during the years of the study. Another 2 ha site served as a reference orchard and was treated in a conventional way. It was located in the same region. Nine to eleven chemical treatments were applied during each season to a conventionally treated reference orchard for control of OFM, PTB and other pests. Five to seven of them were applied for control of OFM and PTB Monitoring of OFM and PTB flights was carried out using sex pheromone trapping in the years of the study. PHEROCON® VI Delta sticky traps were installed in the trial orchard using a scheme provided by the producer. The traps were baited with standard OFM L2 – Orfamone and PTB L2 – Anemone and changed every 8 weeks. We used PHEROCON® OFM COMBO A&B DUAL lures, a new product developed by Trécé Inc., USA for the

PheroFip 2019 177

orchards with MD in the trial orchard in 2017 and 2018. The lures were changed every 8 weeks. The traps were installed before OFM and PTB flights started. For comparison, PHEROCON® VI Delta sticky traps and PHEROCON® OFM COMBO A&B DUAL lures were installed in a reference orchard. All pheromone traps were checked twice per week. The damage of shoots was evaluated during the first generation of OFM and PTB on 20 randomly selected trees, within the central area of each block. Correspondingly, fruit damage was recorded on 100 fruits per each selected tree. Therefore, 2000 fruits were inspected for damage from both pests in each block. The rate of damaged fruits in the trial plots were compared with that in the reference orchard, treated with conventional pesticides only, using the same evaluation techniques. In the reference orchard the fruit damage by OFM ranges from 2.9 to 3.7% and by PTB from 1.2 to 1.4% in the successive years. Percentage of damage in the experimental orchard treated with CIDETRAK® OFM/PTB MESO™ was 0.1% by OFM and 0.1% by PTB i.e. rather below the economical threshold. The significance of differences in the damage rate between the trial and reference orchards was estimated by use of Chi-square tests. The difference in the percentage of damaged fruits between the reference orchard and the trial plots was highly significant for all cultivars under study. Considerable fruit damage was noted in the reference orchard, in spite of many chemical treatments applied. CIDETRAK® OFM/PTB MESO™ is effective when used at a dosage of 20 dispensers per ha, applied once during the season, before the first onset of OFM and PTB. The use of CIDETRAK® OFM/PTB MESO ™ shows that the number of dispensers used does not affect the effectiveness of mating disruption and the reduced rate of dispensers used will help the growers to decrease labor in the field. Applications of these dispensers can provide an effective control of oriental fruit moth and peach twig borer, with better results than the conventional protection programs employed currently in Bulgaria. The CIDETRAK® OFM/PTB MESO™ dispensers, proved to be an effective means of control even in small size orchard lots of one ha. The studies are being continued.

Key words: Cydia molesta Busck, Anarsia lineatella Zell., mating disruption, pheromone dispensers, CIDETRAK® OFM/PTB MESO™

Acknowledgments This study is partially supported by the National Scientific Fund of Bulgaria from the Project No. 16/4, 2017 “COMPETITION FOR FINANCIAL SUPPORT FOR RESEARCH PROJECTS – 2017”

PheroFip 2019 178

Control of plum fruit moth, Grafolitha funebrana Tr., by ISOMATE® - OFM TT dispensers in plum orchards of Bulgaria

D. Stefanova1, H. Kutinkova1, Petar Savov1, R. Andreev 2 ,N. Palagacheva3, M. Tityanov3

1Fruit Growing Institute, Ostromila 12, 4004 Plovdiv, Bulgaria; [email protected] 2Agricultural University, Mendeleev 12, 4000, Plovdiv, Bulgaria 3Summit Agro Romania SRL – Branch Office Bulgaria, 39 Bigla Str., ground floor, office 2, 1164 Sofia, Bulgaria

Abstract Plum is a traditional fruit crop in Bulgaria. The plum fruit moth, Grapholita (syn.Cydia) funebrana (Tr.), is an important and the most difficult pest to control in plum orchards. The larvae of summer generation feed in fruits and caused damage from early summer till the harvest time. For a long time, pest management in stone fruit orchards in Bulgaria relied on organophosphate and pyrethroid insecticides. Considering environmental concerns the eco- friendly means of control, alternative to chemical insecticides are urgently needed. The ecological approach imposes a wider application of the methods of pest management that decrease or completely eliminate use of chemicals polluting the environment within the integrated fruit production systems. Mating disruption (MD) is a promising solution for control of different pests, among them plum fruit moth. The aim of this study was to test the effectiveness of mating disruption (MD) for control of GF (plum fruit moth) in plum orchards using Isomate® - OFM TT dispensers (Shin-Etsu, Japan). The trials were carried out in an isolated 2.5-ha private plum orchard in two consecutive years - 2017 and 2018 Monitoring of GF flight was carried out by sex pheromone trapping in the years of the study. PHEROCON® VI Delta, sticky traps were installed in the trial orchards using a scheme provided by the producer. The traps were baited with standard GF L2 – Funemone lures and changed every 8 weeks. These products - pheromone traps and baits were developed and are manufactured by Trécé Inc., USA. For comparison, 2 standard traps were installed in the reference, conventionally treated orchard located in the same region. All pheromone traps were checked twice a week. During the season, fruit damage was assessed in the trial and reference plot on 1000 fruits each time. At harvest, 1000 fruits were sampled in both orchards, to evaluate the final damage rate. Catches of male moths in pheromone traps were completely inhibited in the MD block, whereas they were numerous in the reference, conventionally treated orchard during both years of our study. Isomate® - OFM TT dispensers, installed before the first flight of Grapholita funebrana males, reduced fruit damage significantly. The percentage of fruits containing plum fruit moth larvae, called red plum maggots, was rather below the Economic Injury Level (EIL)- from 0.5% to 0.6%. In contrast, in the reference, conventionally treated orchard the damage rate was between 7.1 and 9.2 %. The significance of differences in the damage rate between the trial and reference orchards was estimated by use of Chi-square tests. The present study shows that the damage caused by GF larvae in the reference orchard was considerable, in spite of 8 insecticide applications. It is suspected that the population of GF in this orchard had become resistant to some of the insecticides used.

PheroFip 2019 179

In contrast, mating disruption, by means of Isomate® - OFM TT dispensers, ensures an effective protection from plum fruit moth. In view of increasing concern of consumers in internal fruit quality and the regulatory restrictions limiting the use of many insecticides in the EU, this new technology may present a promising alternative to conventional protection programmes in plum orchards of Bulgaria The positive results obtained in this study indicate that mating disruption for control of plum fruit moth may be an effective alternative to conventional (pesticide) treatments. This approach to controlling plum fruit moth is in line with the recent EU recommendations that take care of preservation of the natural environment and production of healthy fruits, with no pesticide residues. The studies are being continued.

Key words: IPM, plum fruit moth, red plum maggot, Grapholitha funebrana, Cydia funebrana, mating disruption, pheromone dispenser, Isomate® - OFM TT

Acknowledgments This study is partially supported by the National Scientific Fund of Bulgaria from the Project No. 16/4, 2017 “COMPETITION FOR FINANCIAL SUPPORT FOR RESEARCH PROJECTS – 2017”

PheroFip 2019 180

Synthesis of San Jose scale sex pheromone and its application in field mating disruption

Takeshi Kinsho1, Akihiro Baba1, Yuki Miyake1, Yasuhiko Kutsuwada1, Tatsuya Fujii2

1Specialty Chemicals Research Center, Shin-Etsu Chemical Co., Ltd., Niigata, Japan 2Fine Chemicals Department, Shin-Etsu Chemical Co., Ltd., Tokyo, Japan

Abstract The San Jose scale (SJS), Quadraspidiotus perniciosus (Comstock), is a cosmopolitan and polyphagous pest, and its infestation makes crops unmarketable, and severe infestations can kill plants. Although the insecticide application against the crawlers in early spring is a well- known conventional control method, it is sometimes insufficient to control the scale, and the later generations are difficult to manage by insecticide spraying. Pheromone-based mating disruption (MD) has been considered as an alternative eco-friendly pest control method of this serious pest. Female produced sex pheromone of SJS consists of three isomeric components, namely, 7-methyl-3-methylene-7-octenyl propionate, (Z)-3,7-dimethyl-2,7-octadienyl propionate and (E)-3,7-dimethyl-2,7-octadienyl propionate. Synthesis employing coupling reaction of the two C-5 building blocks enables the facile construction of the molecule and serves as an industrially scalable method in order to supply sufficient amounts of the active ingredients for actual application in the field. Field mating disruption trials using tube-type dispensers have been conducted in Chile, Argentina and Japan since 2015. MD efficacy was demonstrated through suppression of the male trap capture and reduction of the crawler density.

Key words: San Jose scale, Quadraspidiotus perniciosus (Comstock), pheromone synthesis, mating disruption

PheroFip 2019 181

First results on mating disruption of the vine mealybug in Portugal

Elsa Borges da Silva1,3, Catarina Mourato3, João Vaz3, António Mexia2,3, José Carlos Franco1,3

1CEF, Instituto Superior de Agronomia, Universidade de Lisboa, 1340-017 Lisboa, Portugal; 2LEAF, Instituto Superior de Agronomia, Universidade de Lisboa, 1340-017 Lisboa, Portugal 3Instituto Superior de Agronomia, Universidade de Lisboa, 1340-017 Lisboa, Portugal

Abstract Mealybugs (Hemiptera, Pseudococcidae) are considered economically important pests in vineyards, in different regions worldwide. In a survey conducted in Portugal, on the pest status in vineyards, mealybugs were reported as key pests, occasional pests, or potential pests in 15%, 62%, and 24% of the cases, respectively (Godinho & Franco, 2001). Chemical control is still the most common management tactic used against vine mealybugs. In the same survey, 48% of the responders referred to apply insecticides occasionally to control mealybugs, whereas 28% reported to spray at least once a year. However, chemical control is often ineffective due to mealybug cryptic behaviour, waxy body cover, and clumped spatial distribution pattern. Also, the use of insecticides may be unsustainable, due to resistance, low selectivity, human toxicity, or risk or hazard assessment (Franco et al. 2009). Therefore, there is a need to find sustainable alternative control tactics. Here we present the first results of experiments carried out in Portugal to test the use of mating disruption for pest management of the vine mealybug, Planococcus ficus (Signoret). Mating disruption trials were conducted in three vineyards (Golhelha, Monte do Duque, and Ribafreixo) in the region of Alentejo, in 2017 and 2018. In each vineyard, 2 ha plots of the same grapevine variety were allocated to the studied treatments: mating disruption versus control. In one of the farms (Ribafreixo), the trials were conducted in two 1 ha plots for each treatment. Mating disruption was based on Isonet PF formulation from Shin-Etsu, with a dosage of 500 dispensers per ha, applied early May. Male activity in each plot was monitored weekly, based on captures in four pheromone traps. The level of mealybug infestation in grapevines was estimated in June and July by visual observation of the trunk, branches, and canopy, during 60 s per vine, in a sample of 30 vines per plot, by registering the presence of mealybugs and their stages. The level of infestation in each vine was determined using the following index: 0- no mealybugs; 1 – isolated individuals; 2 – few mealybug colonies; and 3 – several to many colonies. The percentage of infested grape bunches was estimated at harvest, by sampling between 50 (2018) and 100 (2017) bunches per plot, and counting the number of mealybugs per bunch. Mean male captures in pheromone traps exposed within control plots were significantly higher (4.7 times) those in mating disruption treatment. The disruption index varied between 36% and 86%, in 2017, and between 77% and 80%, in 2018, depending on vineyards. The mean level of mealybug infestation in vine canopy during June-July in control plots was about 1.4 times higher but not significantly different from that in mating disruption. However, at harvest, the mean grape infestation, in mating disruption was significantly lower than in control plots. Further experiments will be carried out in 2019.

Key words: Planococcus ficus, mealybugs, grapevine, pheromones, IPM

PheroFip 2019 182

Acknowledgements Thanks are due to Francisco Mata, António Mau, João Maria Correia, Filipe Perdiz, Hugo Pardal, André Pilirito e Andreia Ribeiro from ATEVA, for the help in the selection of the experimental plots, application of mating disruption dispensers, and field monitoring; to João Correia, Miguel Feijão, and Nuno Bicó for allowing us to conduct the experiments in their farms; to Vera Zina, André Garcia, and Mauricio Bigolin, for the help in field sampling; to Biosani and Shin-Etsu, for providing the pheromone dispensers used in the experiments. We also acknowledge the financial support of KKL-JNF Israel (2015–2017) and “Grupo Operacional IntenSusVITI, Intensificação sustentável da vitivinicultura através da poda mecânica, PDR2020-1.0.1-FEADER-03200, Parceria nº 82 / Iniciativa nº 164”. CEF is a research unit funded by FCT (UID/AGR/00239/2013).

References Franco, J. C., Zada, A. & Mendel, Z. 2009: Novel approaches for the management of mealybug pests. In: Biorational Control of Arthropod Pests: Application and Resistance Management (eds. Ishaaya, I. & Horowitz, A.R.): 233–278. Springer, Dordrecht, The Netherlands Godinho, M. A. & Franco, J. C. 2001: Survey on the pest status of mealybugs in Portuguese vineyards. IOBC wprs Bulletin 24(7): 221-226.

PheroFip 2019 183

The potential of pheromones for controlling Pseudococcus calceolariae (Hemiptera: Pseudococcidae) in fruit crops

Tania Zaviezo1*, Jan Bergmann2, Alda Romero1, Ivan Osorio1, Fernanda Flores2, Carolina Ballesteros1

1Departamento Fruticultura y Enología, Facultad Agronomía e Ingeniería Forestal, Pontificia Universidad Católica de Chile, Santiago, CHILE. 2Instituto de Química, Pontificia Universidad Católica de Valparaíso. Avda. Universidad 330, Curauma, Valparaíso, Chile. *[email protected]

Abstract Mealybugs (Hemiptera: Pseudococcidae) are economically important pests of many fruit crops and the pheromones of several species have been identified. For testing its potential use in the control of Pseudococcus calceolariae in fruit crops in Chile, we carried out several field experiments, including the approaches of mating disruption, mass trapping and attract and kill. In mating disruption trials male captures before treatments were similar for control and mating disruption plots, while after captures declined significantly. Nevertheless, significant differences in visual counts were not found and populations were very low in all plots. In the mass trapping trial, plots with pheromones had captures between 36 and 40 times that of plots without pheromones, and sticky panels catches were between 13 and 30 times. No significant differences for visual counts were detected. The attract and kill experiment is underway. Pheromones show a great potential as a direct control tactic for P. calceolariae in fruit crops.

Introduction Mealybugs (Hemiptera: Pseudococcidae) are economically important pests of many fruit crops in several regions of the world, mainly due to their quarantine status for particular markets. The citrophilus mealybug, Pseudococcus calceolariae (Maskell), is a pest thought to be native from Australia and now present in many continents (Williams, 1985; Daane et al., 2012; García Morales et al. 2016). It is a polyphagous insect present in many plant families (García Morales et al. 2016), and responsible for economic losses in fruit crops such as citrus, avocados, grapes, apples, and berries (Ripa & Larral, 2008; Charles et al. 2010; Daane et al. 2012; Dreistadt, 2012). The first sex pheromones of mealybug species were identified in the early 1980’, but it wasn’t until the early 2000’s that many new species were added, including that of P. calceolariae in 2010 (El-Sayed et al. 2010; Unelius et al. 2011). Recently, we have explored the use of P. calceolariae pheromone for its monitoring in fruit crops (Flores et al. 2015). The objective of this work was to evaluate P. calceolariae pheromone as a direct control measure in fruit crops through the approaches of mating disruption, mass trapping and attract and kill.

Material and methods Mating disruption: We have carried out experiments for three seasons in apple orchards and two seasons in citrus orchards. In apples, the first season a low and a high infestation orchards were used. In each orchard six 0.1 ha plots were set: three control (without pheromone) and 3 mating disruption plots. In the second and third season the high infestation apple orchard and a citrus orchard were used. In these two seasons, ten 0.1 ha plots were set in each orchard,

PheroFip 2019 184

with half corresponding to the control treatment and half to the mating disruption treatment. The mating disruption treatment corresponded to 6.2 g of pheromone in a SPLAT formulation, uniformly distributed in the plots. In season one, the mating disruption treatment was set on February, in season two in December and in season three in November. Populations were monitored by pheromones traps and visual inspections to plants, from spring to fall (September to April). Comparisons between control and mating disruptions treatments before and after setting the pheromones in the field were made. Mass trapping: For this experiment an apple and a blueberry orchard were selected. Eight 10 x 10m plots were established in each orchard and in the 4 corners of each plot a trap and a sticky transparent panel (10 x 8 cm aprox.) hanging from it were set. In half of the plots a pheromone dispenser was added and in the other half a dispenser without pheromone. Traps and panels were replaced monthly (October to April) and brought to the laboratory to count males. Populations in the plants at the centre of the plot were monitored visually. Season long male catches and populations were compared between the control and mass trapping treatment. Attract and kill: For this experiment three orchards were selected, one table grape, one citrus and one blueberry, and a combined formulation was prepared using SPLAT. The lure contained the pheromones of P. calceolariae, P. viburni and P. longispinus. Treatments were (1) mix of pheromones plus insecticide, (2) mix of pheromones without insecticide and (3) control (SPLAT alone, no pheromones and no insecticides), with three replicates each. The treatment was set on November. Populations are being monitored by pheromones traps and visually from September to the end of the season. Comparisons between the three treatments before and after setting the pheromones in the field will be compared.

Results and discussion Results of mating disruption trials showed that there was a significant effect of the pheromone treatment in male captures in all orchards and field seasons. Captures before treatments were similar between control and mating disruption plots, while after male captures declined significantly (“trap shutdown”) indicating and important disruption in male orientation (Figure 1). Nevertheless, significant differences in visual counts were not found, in part due to the low popultions present in all plots. In the case of mass trapping, in plots with pheromones trap captures were between 36 and 40 times that of plots without pheromones, and for the sticky panels it was between 13 and 30 times. Here no significant diffrences for visual counts were detected. The attract and kill experiment is underway, and results will be presented in the meeting. This study shows the potential use of pheromones as a direct control tactic for P. calceolariae in fruit crops. Multiyear trials are necessary to fully evaluate its effects. Its final application as a control measure will depend on an economic assessment of the costs, along with the benefit of lower pesticide residues in the product.

PheroFip 2019 185

Figure 1: Males captures in pheromone traps in mating disruption plots (Treated) and control plots (Control), before and after applying the pheromones. Different panels represent different orchards and or seasons.

Key words: mealybugs, mating disruption, mass trapping, attract and kill

Acknowledgements We thank the Grants FONDEF D10i1208 and FIA FIA PYT-2017-0140 from the Chilean government.

References Charles, J. G., Bell, V. A., Lo, P. L., Cole, L. M. & Chhagan, A. 2010: Mealybugs (Hemiptera: Pseudococcidae) and their natural enemies in New Zealand vineyards from 1993–2009. N. Z. Entomol. 33: 84–91. Daane, K. M., Almeida, R. P. P., Bell, V. A., Walker, J. T. S., Botton, M., Fallahzadeh, M., Mani, M., Miano, J. L., Sforza, R., Walton, V. M. & Zaviezo, T. 2012: Biology and management of mealybugs in vineyards, pp. 271–307. In Bostanian, N. J., Vincent, C. & Isaacs, R. (eds.), Arthropod Management in Vineyards: Pests, approaches, and future directions. Springer, New York,NY. Dreistadt, S. 2012: Integrated pest management for citrus, 3rd Edn. University of California Publication Number: 3303. El-Sayed, A., Unelius, R., Twidle, A., Mitchell, V., Manning, L., Cole, L., Suckling, D. M., Flores, M.F., Zaviezo, T. & J. Bergmann. 2010: Chrysanthemyl 2-acetoxy-3- methylbutanoate: the sex pheromone of the citrophilus mealybug, Pseudococcus calceolariae. Tetrahedron Lett. 51: 1075–1078.

PheroFip 2019 186

Flores, M. F., Romero, A., Oyarzún, M. S., Bergmann, J. & Zaviezo, T. 2015: Monitoring Pseudococcus calceolariae (Hemiptera: Pseudococcidae) in fruit crops using pheromone- baited traps. J. Econ. Entomol. 108: 2397–2406. García Morales, M., Denno, B. D., Miller, D. R., Miller, G. L., Ben-Dov, Y., & Hardy, N. B. 2016: ScaleNet: A literature-based model of scale insect biology and systematics. Database. doi: 10.1093/database/bav118. http://scalenet.info. (accessed 20 October 2018). Ripa, R., & Larral. P. 2008: Manejo de Plagas en Paltos y Cítricos. Colección Instituto de Investigaciones Agropecuarias No 23. Centro Regional de Investigación La Cruz, p. 399. Unelius, R., El-Sayed, A., Twidle, A., Bunn, B., Zaviezo, T., Flores, M. F., Bell, V., & Bergmann, J. 2011: The absolute configuration of the sex pheromone of the citrophilus mealybug, Pseudococcus calceolariae. J. Chem. Ecol. 37: 166–172. Williams, D. J. 1985: Australian Mealybugs. British Museum (Natural History), London Publication 953, p. 431.

PheroFip 2019 187

SESSION 5

SIT AND BIOLOGICAL CONTROL OF

FRUIT CROP PESTS

CHAIRPERSON: KERSTIN KRÜGER

PheroFip 2019 188

The Sterile Insect Technique for Mediterranean fruit fly control: a North- Italy pilot project

Serena G. Chiesa1*, Gino Angeli1, Massimo Cristofaro2, Silvia Arnone2, Claudio Ioriatti1

1Fondazione Edmund Mach, via E. Mach 1, 38010 San Michele all’Adige (TN), Italy. 2 Enea - Centro Ricerche Casaccia – Roma. *[email protected]

Abstract The Mediterranean fruit fly (Ceratitis capitata Wiedemann) is a pest of Eastern African origin and was established long time ago in fruit areas of the Mediterranean coast (Balachowski, 1950). It has been described as one of the world's most threatening agricultural pest, attacking more than 300 different host fruit (Liquido et al. 1990, Papadopolus et al. 2001, De Meyer et al. 2002, Malacrida et al. 2007). Historically, it affected the fruit crops of southern and central Italian regions, where it develops on citrus, apricot, peach, fig, apple and khaki. Recently, due to climate changes, fruit damages has been recorded also in the northern regions as Trentino- SouthTyrol (Gutierrez & Ponti, 2011). In Trentino, C. capitata was reported for the first time in 1990 when infested apple were found in an orchard close to a supermarket where rotten fruits were thrown away. This first report was followed by others testifying a slow and constant diffusion of the pest in the region. At present, it is established in scattered areas in the Adige valley, where it causes economic damage specially on apple, the main fruit crop of the area. To maintain the damage under an economic threshold, specific insecticide treatments with ovo-larvicidal insecticides are necessary in some years. Medfly attacks apple during ripening period and insecticide applications to control the infestation potentially could increase pesticide residues on fruit. To avoid pesticide residue issues, research efforts have been focused on the development of environmental friendly approaches to pest management, including mass-trapping, attract and kill and Sterile Insect Technique (SIT) (Navarro‐Llopis, et al. 2013; Martinez‐Ferrer et al. 2012; Hendrichs et al. 2002). The success of these alternative control measures depends on the density and the spatial distribution of the target pest. Since the population level of C. capitata in Trentino is still low and the pest is spread in a circumscribed area bordered by forestry and non-host vegetation, the region seems to be suitable for testing the efficacy of these control strategies. Here, we refer on the preliminary steps made by our research group for implementing SIT as measure to control MFF in apple orchard. In fact, as well as the population level, the effectiveness of SIT is undoubtedly associated with the ability of irradiated males to compete with wild males for mating with wild females (Nikolouli et al. 2018). The released sterile males mate with wild females and the eggs laid by the latest will not be viable and will result in no offspring. The competitiveness of released sterile males might be impacted by the insect strain and the rearing method, radiation sterilization, marking, stress during cold storage, shipment to the release site and release procedure (Dyck et al. 2005). In our case, insects belonging to genetic sexing strain Vienna 8 (obtained from IAEA, Seibersdorf, Austria), have been provided by Bioplanta (Valencia, Spain) by TRAGSA, that produces 500 milion sterile pupae/week, sterilized by exposure to beta radiation. Even if the International Plant Protection Convention (IPPC, 2005) includes the sterile insects produced in biofactory as organic control agents, allowing international

PheroFip 2019 189

exchange and potential use in the crops defense, airfreight and customs clearance are still an issue. Our first purpose was to set up a dynamic procedure for transferring the insects from the biofactory to the field. Quality control bioassays on the insects at the arrival is also an essential issue to be evaluated prior to field applications (Dyck et al. 2005). Hence, the effect of the stress suffered during shipment on emergence rate and adult longevity were assessed. The insects at pupal stage are sent once a week by airplanes to Rome. At the airport, the parcel with an appropriate certification goes through customs clearance and immediately is given to a courier for a fast delivering to Fondazione Edmund Mach (FEM) in San Michele all’Adige (Trentino). The time spent by the pupae under hypoxia is critical and should be minimized, otherwise could affect quality parameters. Once received at FEM, the pupae were divided in batches, two for each release point, and kept in the climatic chamber under controlled conditions (T 24±1°C, RH 70±5 %) and supplied with water and food (water, agar 1% and sugar 18%). Quality control were performed registered the percentage of male emerging for each shipment, that was between 52% and 98%, whit an average of 83%. Flight performance was assessed by releasing the insects in three areas of about 10 hectares each along Adige valley, mainly cultivated as apple orchards. The adult release is performed twice a week from defined points, one/ha, from the beginning of July until the end of November. Two types of trap are used to evaluate the effectiveness of the SIT strategy and to monitoring the distribution of sterile male flies within the released areas: Delta sticky trap (Biogard Delta trap - Biogard, CBC) activated with Trimedlure (ISAGRO) attractive for males, and Vaso-trap® with glass jar (by Tap-trap) baited with food attractive (Unipack® Biolure, Suterra) in order to capture both males and females. The results indicate that the sterile males, in our environment, move more than 300 m in every direction and are evenly distributed in the release areas.

Key words: Ceratitis Capitata, Medfly, Sterile Insect Techniques, Trentino, Italy, apple

References (FAO) Food and Agriculture Organization of the United Nations. 2005b: Glossary of phytosanitary terms. ISPM/NIMP/NIMF, Publication Number 5, International Plant Protection Convention (IPPC). FAO, Rome, Italy. Balachowski, M. 1950: Sur l’origine de la Mouche de fruits (Ceratitis capitata Wied.). C R Acad Agric Fr 36: 259–362. De Meyer, M., Copeland, R. S., Lux, S. A., Mansell, M., Quilici, S., Wharton, R. A., White, I. M. & Zenz, N. 2002: Annotated checklist of host plants for Afrotropical fruit flies (Diptera: Tephritidae) of the genus Ceratitis. Documentations zoologiques Vol. 27, Royal Museum for Central Africa, Tervuren, Belgium. Dyck, V. A, Hendrichs, J & Robinson, A. S. 2005: Sterile insect technique: principles and practice in area-wide integrated pest management. Springer, The Netherlands, 787. (FAO) Food and Agriculture Organization of the United Nations 2007: Guidance for packing, shipping, holding and release of sterile flies in area-wide fruit fly control programmes. FAO, Rome 2007. Gutierrez, A. P., & Ponti, L. 2011: Assessing the invasive potential of Mediterranean fruit fly in California and Italy. Biological Invasions, Vol 3, 12: 2661-2676.

PheroFip 2019 190

Hendrichs, J., Robinson, A. S., Cayol, J. P. & Enkerlin, W. 2002: Medfly areawide sterile insect technique programmes for prevention, suppression or eradication: the importance of mating behavior studies. Florida Entomologist 85(1): 1-13. Liquido, N. J., Cunningham, R. T. & Nakagawa, S. 1990: Host plants of Mediterranean fruit- fly (Diptera, Tephritidae) on the island of Hawaii (1949-1985 Survey). J. Econ. Entomol. 83: 1863-1878. Malacrida, A. R., Gomulski, L. M., Bonizzoni, M., Bertin, S., Gasperi, G. & Guglielmino, C. R., 2007: Globalization and fruit fly invasion and expansion: the medfly paradigm. Genetica 131: 1–9. Martinez‐Ferrer, M. T., Campos, J. M., & Fibla, J. M. 2012: Field efficacy of Ceratitis capitata (Diptera: Tephritidae) mass trapping technique on clementine groves in Spain. Journal of applied entomology, 136(3): 181-190. Navarro‐Llopis, V., Primo, J. & Vacas, S. 2013: Efficacy of attract‐and‐kill devices for the control of Ceratitis capitata. Pest management science 69(4): 478-482. Nikolouli, K., Colinet, H., Renault, D., Enriquez, T., Mouton, L., Gibert, P., Sassu F., Cáceres C., Stauffer C., Pereira R & Bourtzis, K. 2018: Sterile insect technique and Wolbachia symbiosis as potential tools for the control of the invasive species Drosophila suzukii. Journal of pest science 91: 489–503. Papadopoulos, N. T., Katsoyannos, B. I., Carey, J. R. & Kouloussis, N. A. 2001: Seasonal and annual occurrence of the Mediterranean fruit fly (Diptera: Tephritidae) in northern Greece. Ann. Entomol. Soc. Am. 94: 41-50.

PheroFip 2019 191

Genetic diversity and initial population size affect the early colonization success of Mastrus ridens, a parasitoid used for the control of codling moth

Tania Zaviezo1*, Carlos Brochero1, Sofía Miranda1, Thibaut Malausa2

1Departamento Fruticultura y Enología, Facultad Agronomía e Ingeniería Forestal, Pontificia Universidad Católica de Chile, Santiago, CHILE. 2INRA, CNRS, Université Côte d'Azur, ISA, FRANCE *[email protected]

Abstract Mastrus ridens Horstmann (Hymenoptera: Ichneumonidae) is one of the main parasitoid species attacking codling moth (Cydia pomonella [L.]) in its region of origin (southern Kazakhstan) (Mills, 2005; Retamal et al. 2016). It has been used in several classical biological control programs against codling moth in North and South America, New Zealand, Australia, and is being considered for release in Europe (Mills, 2005; Devotto et al. 2010; Sandanayaka et al. 2011; Tortosa et al. 2014). The biology of Mastrus ridens correspond to a specialist parasitoid that attacks mature codling moth larvae in their cocoons, by paralyzing and then laying several eggs on the cuticle of its host, i.e. it corresponds to a gregarious ectoparasitoid. As most hymenopterans, this species has a haplodiploid mode of reproduction, but additionally it displays Complementary Sex Determination (CSD) (Retamal et al. 2016). As a result, populations with low genetic diversity or under inbreeding conditions produce less female progeny and larger proportion of diploid males (Retamal et al. 2016; Zaviezo et al. 2017). The aim of this study was to examine the effect of initial genetic diversity and founding numbers on M. ridens population growth, and other associated parameters, in laboratory conditions. For testing the effect of genetic diversity, we created M. ridens low diversity (isofemale lines) and high diversity lines (mix of several lines). Then six mated females were put inside a plastic box (19.5 x 13 x 9 cm) with codling moth larvae and honey, creating in this way treatments with the same initial population numbers, but different genetic diversity depending on the line used in each box. From then on, additional larvae were added periodically, and at day 21 and 42 a new box was added on top, connected through tubes. Population was estimated every other day by observing and counting the insects trough the box walls. At day 70 the experiment ended, and all individuals present in the boxes were counted, including codling moth larvae and adults. For testing the effect of founding numbers, we created more complex arenas made of plastic boxes (23 x 13 x 4,5 cm) with ten internal divisions conected by one small perforation on each division. In each box, either 1 unmated female and 1 male was added or 4 unmated females and 4 males. All insects within a box belonged to the same low diversity line, but different lines were used in the boxes. In this way, we varied initial numbers but the level of genetic diversity was low in all cases. As in the previous experiments, codling moth larvae were added periodically, and also new boxes above the initial ones as time passed. At day 45 the experiment ended and all individuals present in the boxes were counted, including codling moth larvae and adults. Results showed that there was an effect of genetic diversity in population growth being 17.5 and 11.0 for the whole period in the high and low diversity treatments respectively (p = 0.047). High diversity treatments generated 1.6 times the number of individuls than low diversity treatments. These diferences were even larger and significant when comparing the

PheroFip 2019 192

number of females produced, which was twice as much for the high diversity treatment (p = 0.04). Also the overall sex ratio (proportion of males) was lower in the high diversity treatment (0.51 versus 0.61, p = 0.02). We also found an effect of initial population numbers. The most important being establishment rate (proportion of boxes where production of female offspring occurred), which was 44% in the low population treatment and 83% in the high population treatment. Also, as expected, total numbers of individuals produced, and total number of females was larger in the high population treatment (p < 0.001 for both). This study shows that both, genetic diversity and populations numbers affect population growth, female production and sex ratio, and thus early colonization success of these experimental laboratory populations. These same effects could be taking place in the field, when low numbers are released or laboratory colonies are initiated, and thus this should be considered when carrying out classical or augmentative biological control programs.

Key words: biological control, sex ratio, apple

Acknowledgements We thank the Chilean government for funding through FONDECY program, projects # 1131145 and 1181256.

References Mills, N. J. 2005: Selecting effective parasitoids for biological control introductions: Codling moth as a case study. Biol. Control 34: 274-282. Devotto, L., del Valle, C., Ceballos, R., & Gerding, M. 2010: Biology of Mastrus ridibundus (Gravenhorst), a potential biological control agent for area-wide management of Cydia pomonella (Linneaus) (Lepidoptera: Tortricidae). J. Appl. Entomol. 134: 243-250. Retamal, R., Zaviezo, T., Malausa, T., Fauvergue, X., Le Goff, I., & Toleubayev, K. 2016: Genetic analyses and occurrence of diploid males in field and laboratory populations of Mastrus ridens (Hymenoptera: Ichneumonidae), a parasitoid of the codling moth. Biol. Control 101: 69–77. Sandanayaka, W. R. M., Chhagan, A., Page-Weir, N. E. M., & Charles, J. G. 2011: Colony optimization of Mastrus ridens (Hymenoptera: Ichneumonidae), a potential biological control agent of codling moth in New Zealand. New Zeal. Plant Prot. 64: 227-234. Tortosa, O. E., Carmona, A., Martínez, E., Manzano, P., & Giardina, M. 2014: Liberación y establecimiento de Mastrus ridens (Hymenoptera: Ichneumonidae) para el control de Cydia pomonella (Lepidoptera: Tortricidae) en Mendoza Argentina. Rev. Soc. Entomol. Argentina 73:109-118. Zaviezo, T., Retamal, R., Urvois, T., Fauvergue, X., Blin, A., & Malausa, T. 2018: Effects of inbreeding on a gregarious parasitoid wasp with complementary sex determination. Evol. Appl. 11: 243–253.

PheroFip 2019 193

Parasitization by Aphelinus mali on Eriosoma lanigerum: an IPM and Organic comparison

Serena Giorgia Chiesa1*, Luca Corradini1, Mario Baldessari1, Gino Angeli1

1Fondazione Edmund Mach, Technology transfer center, Via E. Mach 1. I-38010 - San Michele all’Adige (TN) Italy *[email protected]

Abstract The woolly apple aphid (WAA), Eriosoma lanigerum (Hausmann), is a pest of apple orchards infesting both stems and roots of host trees (Baker, 1915). Heavy infestations of WAA can cause yield and growth reduction of apple trees, as this aphid weakens the plant by both feeding on roots and inducing root and shoot galls that damage the xylem and interrupt water transportation (Klimstra & Rock, 1985), and even death of young trees (Brown & Schmitt, 1990; Brown et al. 1994). The pest status of WAA is fluctuating over the years depending on biotic and abiotic factors (Lordan et al. 2015; Angeli, 2015). The control of the woolly apple aphid by the solitary, host-specific parasitoid Aphelinus mali (Haldeman) (Hymenoptera: Aphelinidae) is cited as one of the most successful examples of classical biological control of insect pests (Mueller et al. 1992; Goossens et al. 2011). De Bach (1964) reported A. mali as a successful biological control agent in at least 40 countries and among these also Italy. The susceptibility of some apple varieties and dwarfing rootstocks, the simplification of the agroecosystems and the management programs of primary pests and diseases are reported as potentially responsible for disrupting the biological control of WAA (Angeli & Simoni, 2006; Baldessari et al. 2007; Beers, 2010; 2014; Angeli, 2015). Some recently introduced agronomic techniques, such as single row exclusion netting, are also negatively involved, reducing the possibility of colonization of the plant's foliage by predators and parasitoids. E. lanigerum becomes active starting from 267 degree-days above the temperature threshold of 5.2°C (Asante et al. 1991), which generally corresponds to the crop stage of green bud (BBCH 56), whereas A. mali, whose lowest developmental threshold is 8.3°C, needs 255 degree-days to begin its activity (Asante & Danthanarayana, 1992). Consequently, the developmental time of A. mali is generally delayed throughout the year compared to its aphid host. A two-year field experiment (2017-18) was conducted in some organic and IPM orchards (Golden Delicious and Fuji varieties) located in plain (180 m) and hilly areas (500-700 m) of Trentino-South Tyrol Region (north-eastern Italy) to determine the efficacy of a biological control of WAA populations by means of Aphelinus mali. The evolution of plant colonization by WAA, the flight of the parasitoid A. mali and the percentage of parasitization of WAA colonies were monitored in the selected orchards. A particular attention was paid at evaluating the susceptibility of A. mali to the common pesticides used in the two crop management systems and in relation to varieties and altitude of cultivation. Our results suggest that during summer the parasitoid A. mali has certainly an impact on WAA, but insights on the single interactions do not indicate a sufficient biocontrol in the period May‐half June, both in IPM and in organic management, in plain and in hilly areas, and also considering the different apple varieties. It was confirmed (Goossens et al. 2011) that the insufficient contribution of the early parasitation by A. mali to avoid woolly apple aphid damage is linked to a later emergence

PheroFip 2019 194

from diapause and a slower reproduction compared to its host. A delay of parasitoid development in hilly areas has been observed. Noteworthy, observing the first peak of flight of A. mali in the post-flowering period (6th- 20th June), although limited in size, is essential for an exponential flight increase and the consequent culmination of the parasitization ratio in summer, generally at the end of July. This constitutes a valuable complement in the integrated control strategy of E. lanigerum. The monitoring of the number of black mummies parasitized and of the A. mali adults’ flight in the four orchards along the season allowed also to evaluate the development of endoparasitic stages, which covers the same period in the studied orchards, delayed of about one week on the hills. This phase, in which A. mali larvae reside inside their host, is strategic to position treatments with compounds with a risk of toxicity on A. mali adults, such as some insecticides, to obtain a better selectivity. Outbreaks of E. lanigerum have often occurred as a result of pesticide applications which decimated biological control agents (McLeod, 1954; Penam & Chapman, 1980). In presence of high infestations of woolly apple aphid, the chemical control of D. plantaginea with spirotetramat (+ oil) or other aphicides, which remains necessary in IPM, shows a favorable side-effect on WAA in order to anticipate the high migration waves and the subsequent colonization of growing shoots. In this two-year work, there were no differences in the parasite-host relationships in the two crop management systems, IPM and organic, even though a higher presence of WAA was observed in organic orchards. The climatic conditions seem to play a key role in regulating the ability of A. mali to control aphid infestations. Investigations are still going on, especially to set up the optimal management of parasitoids through simple practices, such as mowing at alternate rows.

Key words: monitoring, Aphelinus mali Haldeman, parasitoids, Eriosoma lanigerum Hausmann, wolly apple aphid, IPM, organic.

References Angeli, G. 2015: Nuove prospettive di difesa dagli afidi del melo. L’Informatore Agrario 13: 50-51. Angeli, G. & Simoni, S. 2006: Apple cultivars acceptance by Dysaphis plantaginea Passerini (Homoptera: Aphididae). J. Pest Sci. 79: 175-179. Asante, S. K., Danthanaryana, W. & Heatwole, H. 1991: Bionomics and population growth statistics of apterous virginoparae of woolly apple aphid, Eriosoma lanigerum, at constant temperatures, Entomol. Exp. Appl. 60: 261-270. Asante, S. K. & Danthanarayana, W. 1992: Development of Aphelinus mali an endoparasitoid of woolly apple aphid, Eriosoma lanigerum at different temperatures. Entomol. Exp. Appl. 65(1): 31-37. Baldessari, M., Rizzi, C., & Angeli, G. 2007: La difesa dell'afide lanigero sul melo in Trentino. L'informatore agrario 63: 20-22. Beers, E. H., Cockfield, S. D. & Gontijo, L. M. 2010: Seasonal phenology of woolly apple aphid (Hemiptera: Aphididae) in Central Washington. Environ. Entomol. 39(2): 286- 294. Beers, E. H. 2014: Wolly Apple Aphid. In T.B. Sutton, H. S. Aldwinckle, A.M. Agnello and J.F. Walgenbach (eds), Compendium of Apple and Pear Diseases and Pests, Second Edition, APS Press. 174-175. DeBach, P. 1964: Biological Control of Insect Pests and Weeds, Reinhold Publishing Corporation, New York.

PheroFip 2019 195

Goossens, D., Bangels, E., Belien, T., Schoevaerts C. & De Maeyer L. 2011: Optimal profit of the parasitation by Aphelinus mali in an IPM complementary strategy for the control of Eriosoma lanigerum. Commun. Agric. Appl. Biol. Sci. 76(3): 457-465. Lordan, J., Alegre, S., Gatius, F., Sarasúa, M. J. & Alins, G. 2015: Woolly apple aphid Eriosoma lanigerum Hausmann ecology and its relationship with climatic variables and natural enemies in Mediterranean areas. Bull. Entomol. Res. 105(1): 60-69. Mueller, T. F., Blommers, L. H. M. & Mols, P. J. M. 1992: Woolly apple aphid (Eriosoma lanigerum Hausm., Hom., Aphidae) parasitism by Aphelinus mali Hal. (Hym., Aphelinidae) in relation to host stage and host colony size, shape and location. J. Appl. Entomol. 114: 143-154.

PheroFip 2019 196

SESSION 6

APPLIED BIOTREMOLOGY: A NEW

DISCIPLINE FOR PEST MANAGEMENT.

VIBRATIONAL SIGNALS AS

SEMIOPHYSICALS

CHAIRPERSON: VALERIO MAZZONI

PheroFip 2019 197

Evaluation of the social behavior based on vibrational signals of Philaenus spumarius in semi-field conditions

Imane Akassou1,2, Sabina Avosani1,2, Vincenzo Verrastro3, Valerio Mazzoni2

1DICAM Department of Civil, Environmental and Mechanical Engineering, University of Trento, Trento, Italy; 2Research and Innovation Centre, Fondazione Edmund Mach, San Michele all'Adige (TN), Italy; 3CIHEAM–IAMB - International Centre for Advanced Mediterranean Agronomic Studies, Bari, Italy

Abstract Substrate-borne vibrations are probably the most common means of communication in insects; they underlie insect social and ecological interactions in a complex vibrational environment, containing interferences from other species and sources of noise. In leafhoppers (Hemiptera: Cicadomorpha), the mating behavior is mediated by vibrational signals that are emitted in different social contexts: individuals, pairs, and groups (Tishechkin, 2003). The meadow spittlebug Philaenus spumarius (Hemiptera: Aphrophoridae) is the main vector of the quarantine bacterium Xylella fastidiosa, that is the causal agent of the Olive Quick Decline Syndrome, a severe vascular disease that appeared suddenly few years ago in the province of Lecce (Salento peninsula, southeastern Italy) (Saponari et al. 2014). Since the nymphal stage of P. spumarius commonly lives in spittles, grouped on the same plant or on nearby plants; sharing the same feeding site. It is relevant to assess if this aggregation behavior is maintained also in the adult stage and, if it is based on the exchange of vibrational signals. Furthermore, the control of the insect relies on chemicals; it would be interesting to evaluate the potential development of a sustainable management strategy, based on vibrational signals, as physical disruption (Polajnar et al. 2016). Thus, it is fundamental to understand the temporal and the seasonal pattern of the emitted vibrational signals. The aim of our study was to investigate the social behavior, based on vibrational communication signals, between individuals of the same sex of P. spumarius and to evaluate the insect vibrational signaling activity throughout the day but also during the adults development stages. To minimize the communication associated to the mating behavior, the studied groups consisted on 6 to 10 individuals, of only males and only females. They were placed on grapevine plants, separately in two net-cages. Recordings were carried out outdoor, in semi- field conditions, simultaneously via two laser Doppler vibrometers. Trials took place from the middle of June until the end of September, in terms of periods of 20 to 30 days each, in order to evaluate the seasonal pattern of the insect vibrational signaling activity. To study the temporal pattern, three slots of the day were chosen to record: from 06:30 am to 11:00 am, from 11:30 am to 16:00 pm and from 16:30 pm to 21: 00 pm, each slot was repeated 3 times during each period. Our preliminary results indicate that males were calling throughout the full season of recording; calls were usually followed by jumping of the males from the plants (call & fly behavior), while females started calling only later, from the middle of July. During the day; females were emitting more calling signals in the evening, wheras males were emitting calling signals during the three slots of the day, with a lower activity in the middle of the day. The acquired knowledge is important to better understand the signaling behavior and the phenology of P. spumarius in natural conditions. The next steps will consist on the characterization of the recorded signals and their association to their behavioral meanings.

PheroFip 2019 198

Further experiments applying playbacks, in different social contexts (males & females) will be performed, to achieve these goals.

Key words: Philaenus spumarius, vibrational communication, social behavior, substrate- borne signal

References Polajnar, J., Eriksson, A., Virant-Doberlet, M., & Mazzoni, V. 2016: Mating disruption of a grapevine pest using mechanical vibrations: from laboratory to the field. Journal of pest science 89(4): 909-921. Saponari, M., Loconsole, G., Cornara, D., Yokomi, R. K., De Stradis, A., Boscia, D. & Porcelli, F. 2014: Infectivity and transmission of Xylella fastidiosa by Philaenus spumarius (Hemiptera: Aphrophoridae) in Apulia, Italy. Journal of economic entomology 107(4): 1316-1319. Tishechkin, D. Y. 2003: Vibrational communication in Cercopoidea and Fulgoroidea (Homoptera: Cicadina) with notes on classification of higher taxa. Russian Entomological Journal 12(2): 127-181.

PheroFip 2019 199

SESSION 7

INVASIVE PESTS: A CHALLENGE FOR IPM

A - BROWN MARMORATED STINK BUG AND

OTHER INVASIVE HEMIPTERANS

CHAIRPERSON: GREG KRAWCZYK

PheroFip 2019 200

Establishment and current status of Halyomorpha halys damaging peaches and olives in the prefecture of Imathia in Northern Greece

Petros Damos1, Polyxeni Soulopoulou2, Thomas Thomidis2

1Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece; 2Alexander Technological and Educational Institute of Thessaloniki, Department of Agricultural Technology

Abstract Halyomorpha halys (Stål, 1855) (Heteroptera: Pentatomidae), or brown marmorated stink bug, is a temperate/subtropical species native to China, Japan, Korea and Taiwan. However, in the last years, the species was detected in different parts of the world, mainly in North America and most of Europe territory (Leskey & Nielsen, 2018, Cianferoni, 2018). In northern Europe, particularly, the species has been recorded in northeastern France in 2012 (Callot & Brua, 2013) it was reported also in Île-de-France (Garrouste et al., 2015, Maurel et al. 2016) and in Corsica (Kriticos et al. 2017). The species was found in 2007 in Liguria, Italy (Maistrello & Dioli, 2014; Dioli et al., 2016) and in 2011 in Athens, Greece (Milonas & Partsinevelos, 2014). H. halys has a wide host range, including fruit trees, field crops, forest trees as well as wild hosts (Bergman et al., 2016). The species feeding may occur on stems, leaves, shoots although nymphs and adults prefer to feed on developing and mature fruits and seeds of their host plan (Martinson et al. 2015). In the current study are presented data regarding the appearance and the damage caused by the species for the first time in fruit trees in Northern Greece. Fieldwork was conducted during 2018 from July to August in two peach fruit orchard plantations located in Nea Nikomidia and Kavasila and one olive tree orchard located in Kavasila in Northern Greece. Each peach orchard, used as an experimental block, had a parallelogram shape and was separated from other orchards by 10-20m wide cultivation road. Each block consisted of about 40 peach trees (industrial candy variety Loadel) planted in a regular rectangular 5x5 m grid with 10 rows and 10 columns. The tree height was ca. 2.5–3.5 m, and the trees were ca. 10+ years old. The olive orchards consisted of about 30 olive trees (halkidiki variety) planted in a 5x6 grid with 8 rows and 10 columns. The tree height of was ca. 1.5-2.5 m. No insecticides were applied to these orchards and only matting disruption dispensers were used for the peach orchards against Anarsia lineatella and Grapholita molesta. One 2-year branch, ca 1-2 m above the ground where the majority of fruits, or olives, hang was used as sampling unit. In order to have a representative grid of samples in each experimental block every second tree from every second row of each experimental block was used to collect data. Exhaustive counts of sampling units were performed and all fruits on the given branch were inspected for the presence of Halyomorpha halys individuals as well as feeding damages. In total 132 statistical sampling units were used in this study with all its fruits or olives inspected. The presence of H. halys individuals (mostly nymphs) was very high in both peach and olive orchards inspected. The feeding damage is in most cases sever and results in significant economic lost. The initial damage caused by H. halys caused fruit deformation which in most cases leaded to suberifications. In the peach orchards the most severe damage tended to be located on the periphery of the orchards. The percentage of damage to peaches due to H. halys feeding was higher than 50 percent (63±3%). The percentage of damage with at least one feeding entrance to olives was higher than 80 percent (87±6%). However, currently it is

PheroFip 2019 201

unknown whether population of H. halys may cause sever damages in other fruit orchards which received pesticides. Nevertheless, since the first record of Halyomorpha halys in 2011 in Athens, at present the pest has occurred regularly in Northern Greece causing considerable damage in peaches and olives that were screened. Experience in the USA and Italy has shown that despite being polyphagous this pest has a preference for peach as favourite host (Bariselli et al. 2016). However, in this work the damage occurred in olive trees was significantly higher compared to peach fruits arousing considerable concern. The increasing spread of this pest, as well as its preference in peach and olives, makes urgent need to develop efficient and sustainable management strategies in the near future.

Key words: marmorated sting bug, sampling, integrated control

References Bariselli, M., Bugiani, R. & Maistrello, L. 2016: Distribution and damagecaused by Halyomorpha hakys in Italy. Bulletin OEPP/EPPO Bulletin 0(0): 1–3 Bergmann, E. J., Venugopal, P. D., Martinson, H. M., Raupp, M. J. & Shrewsbury, P. M. 2016: Host plant use by the invasive Halyomorpha halys (Stål) on woody ornamental trees and shrubs. PLoS One 11(2): e0149975. doi.org/10.1371/journal.pone.0149975 Callot, H. & Brua, C. 2013: Halyomorpha halys (Stål, 1855), la Punaise diabolique, nouvelle espèce pour la faune de France (Heteroptera Pentatomidae). L’Entomologiste 69(2): 69– 71. Cianferoni, F., Graziani, F., Dioli, P. & Ceccolini, F. 2018: Review of the occurrence of Halyomorpha halys (Hemiptera: Heteroptera: Pentatomidae) in Italy, with an update of its European and World distribution. Biologia doi.org/10.2478/s11756-018-0067-9 Dioli, P., Leo, P. & Maistrello, L. 2016: Prime segnalazioni in Spagna e in Sardegna della specie aliena Halyomorpha halys (Stål, 1855) e note sulla sua distribuzione in Europa (Hemiptera, Pentatomidae). Rev gad Entom 7(1): 539–548. Garrouste, R., Nel, P., Nel, A., Horellou, A. & Pluot-Sigwalt, D. 2015: Halyomorpha halys (Stål 1855) en Île de France (Hemiptera: Pentatomidae: Pentatominae): surveillons la punaise diabolique. Ann Soc Entomol Fr (NS) 50(3–4): 257–259. Kriticos, D. J., Kean, J. M., Phillips, C. B., Senay, S. D., Acosta, H. & Haye, T. 2017: The potential global distribution of the brown marmorated stink bug, Halyomorpha halys, a critical threat to plant biosecurity. J Pest Sci 90: 1033–1043. Leskey, T. C. & Nielsen, A. L. 2018: Impact of the invasive brown marmorated stink bug in North America and Europe: history, biology, ecology, and management. Annu Rev Entomol 63: 599–618. Maistrello, L. & Dioli, P. 2014: Halyomorpha halys Stål 1855, trovata per la prima volta nelle Alpi centrali italiane (Insecta: Heteroptera: Pentatomidae). Il Naturalista valtellinese 25: 51–57. Martinson, H. M., Venugopal, P. D., Bergmann, E. J., Shrewsbury, P. M. & Raupp, M. J. 2015: Fruit availability influences the seasonal abundance of invasive stink bugs in ornamental tree nurseries. J Pest Sci 88: 461– 468. Maurel, J.-P., Blaye, G., Valladares, L., Roinel, É. & Cochard, P.O. 2016: Halyomorpha halys (Stål, 1855), la punaise diabolique en France, à Toulouse (Heteroptera; Pentatomidae). Carnets Natures 3: 21–25. Milonas, P. G. & Partsinevelos, P. K. 2014: First report of brown marmorated stink bug Halyomorpha halys Stål (Hemiptera: Pentatomidae) in Greece. Bull OEPP/EPPO 44:183–186.

PheroFip 2019 202

Pheromone traps for Halyomorpha halys: a three year comparison in pear orchards

Giacomo Vaccari¹, Stefano Caruso¹, Alberto Pozzebon², Lara Maistrello³

1Consorzio Fitosanitario Provinciale Modena. Via Santi 14, 41123 Modena, Italy; 2Dept. of Agronomy, Food, Natural Resources, Animals and Environment, University of Padova. Agripolis, Viale dell'Università 16, 35020, Legnaro (Pd) Italy;3Dept. of Life Sciences, University of Modena and Reggio Emilia, Via G. Amendola 2, 41122, Reggio Emilia, Italy

Abstract The invasive Halyomorpha halys (Stål, 1855) (Heteroptera, Pentatomidae) is one of the emerging pests for Italian agriculture. First recorded in Modena (northern Italy) in 2012, it spread progressively in Italy with great rapidity (Maistrello et al. 2018). Since 2014, damage on pear, apple, peach and kiwi attributed to this insect were reported in fruit-growing areas of northern Italy (Bariselli et al. 2016). A survey conducted in 2016 by the Technical Office of Cooperative Fruit Modena Group, on 60 farms managing a total of 400 ha of pear orchards in Modena Province, estimated the economic damage induced by H. halys at € 3,072,797 (Maistrello et al. 2017). H. halys has a high dispersal ability and a marked tendency to move between different host plants (Lee & Leskey, 2015). These ethological characteristics make active monitoring techniques of (frappage, and visual inspections) unsuitable for this pest. These difficulties coupled with the strong concern for the high damaging potential of H. halys, led farmers to intensify treatments with broad-spectrum insecticides (Maistrello et al. 2017). For this reason, the identification of traps able to reliably estimate the presence and the abundance of the pest is a fundamental step to re-establish the principles of integrated pest management, reducing the use of insecticides and guaranteeing at the same time a more effective and rational control of H. halys populations. Traps based on aggregation pheromones are currently the most promising tools for monitoring the presence of H. halys and, in the United States, these traps were suggested to support management decision in apple orchards (Short et al. 2017). In previous trials in the provinces of Modena and Reggio Emilia, the Rescue Pheromone Traps proved to be effective in capturing H. halys specimens during the entire production season (Maistrello et al. 2016). Nevertheless, two major critical points were detected, that pose a limitation for their use in commercial fruit orchards. First: a strong variability in catches, probably caused by several external factors such as the attractiveness of surrounding plants, the vigor and the bearing of the plant on which they are installed and the insect population density. Second: an increase of fruit damage on plants located up to 8 meters from the installation point (Vaccari et al. 2018). The aim of this work, carried out in 2016-2018, was to identify the most reliable system for monitoring H. halys in pear orchards, assessing the effectiveness of various trap designs (Rescue Pyramid Trap, AgBio Pyramid Trap, Trécé Double Funnel, Trécé Sticky Sheet) and bait types (Rescue, AgBio, Trécé) available on the Italian market, over the entire production season (beginning of May-second half of October). Every year the trap comparison was replicated four times, and each replicate was located on the external row of an organic/untreated pear orchard, that was bordered by a hedge or an uncultivated area. Within each replicate, traps were spaced every 30-40 m and their position was changed randomly every week. The traps were checked weekly, counting H. halys adults

PheroFip 2019 203

and nymphs. Data were analyzed by means of analysis of the variance for repeated measures. The effect of the thesis was evaluated by F test (α = 0.05) and a t test (α = 0.05) was applied for comparison between the thesis. Preliminary results for 2016-17 show significant differences between trap types and among pheromone lures. Among the commercial combinations (Trap+Bait of the same company) Rescue Trap, proved to be the best devices for monitoring H. halys in pear orchards, provided it is correctly installed on the plant, firmly anchoring the base of the trap to the trunk or to a main branch. Considering only the bait effectiveness, Trécé lures appeared much more effective and reliable than Rescue and AgBio lures. The combination of Trécé lures with pyramid models (either Rescue or AgBio) provided better reliability of the traps in pear orchards.

Key words: brown marmorated stink bug, aggregation pheromone, invasive species, integrated pest management

Acknowledgements This study was funded by the Emilia Romagna region within the Rural Development Plan 2014-2020 Op. 16.1.01- GO PEI-Agri - FA 4B, Pr. «HALYS», coordinated by CRPV.

References Bariselli, M., Bugiani, R. & Maistrello, L., 2016: Distribution and damage caused by Halyomorpha halys in Italy. EPPO Bulletin 46 (2): 332–334. Maistrello, L., Vaccari, G. et al. 2016: Monitoraggio in campo e danni della cimice aliena Halyomorpha halys in Emilia Romagna: da minaccia a problema concreto. In Atti delle Giornate Fitopatologiche, pp. 171–178. Giornate Fitopatologiche, Bologna: CLUEB. Maistrello, L., Vaccari, G., Caruso, S., Costi, E., Bortolini, S., Macavei, L., Foca, G., Ulrici, A., Bortolotti, P. P., Nannini, R., Casoli, L., Fornaciari, M., Mazzoli, G. L. & Dioli P., 2017: Monitoring of the invasive Halyomorpha halys, a new key pest of fruit orchards in northern Italy. Journal of Pest Science 90(4): 1231-1244. Maistrello, L., Dioli, P., Dutto, M., Volani, S., Pasquali, S. & Gilioli, G., 2018: Tracking the spread of sneaking aliens by integrating crowdsourcing and spatial modelling: the Italian invasion of Halyomorpha halys. BioScience, in press Lee, D. H. & Leskey, T. C. 2015: Flight behavior of foraging and overwintering brown marmorated stinkbug, Halyomorpha halys (Hemiptera: Pentatomidae). Bulletin of Entomological Research 105(5): 566-73. Short, B. D., Khrimian, A. & Leskey, T. C. 2017: Pheromone-based decision support tools for management of Halyomorpha halys in apple orchards: development of a trap-based treatment threshold. Journal of Pest Science 90(4): 1191-1204. Vaccari, G., Caruso, S., Pozzebon, A. & Maistrello, L. 2018: Pheromone traps for Halyomorpha halys monitoring: evaluation of side-effects in fruit orchards. In: Book of Abstracts, ECE 2018 - XIth European Congress of Entomology. Napoli, Italy, July 2-6 2018. p. 167.

PheroFip 2019 204

Managing Halyomorpha halys by means of exclusion netting: trials 2016- 2018

Stefano Caruso¹, Stefano Vergnani², Giacomo Vaccari¹, Lara Maistrello³

¹Consorzio Fitosanitario Provinciale Modena. Via Santi 14, 41123 Modena, Italy; ²Cooperativa Orogel. Via Nuova 12/A. Cento di Ferrara, Italy; ³Dipartimento Scienze della Vita, Università di Modena-Reggio Emilia. Via G. Amendola 2, 42122 Reggio Emilia, Italy

Abstract The invasive polyphagous crop pest Halyomorpha halys, also known as brown marmorated stink bug (BMSB), is rapidly spreading in Italy and other European countries. In order to cope with its high harmfulness in fruit orchards, in Emilia-Romagna Region, Northern Italy, farmers have increased the use of broad-spectrum insecticides, resulting in disruption of the most innovative IPM strategies, such as mating disruption (MD) or the use of microbiological products (e.g. CpGV), with serious agro-environmental risks (Maistrello et.al. 2017). Several investigations are underway in order to identify more sustainable control strategies against BMSB that include “IPM-CPR” (Blaauw et al. 2014), “attract and kill”, trap crop (Morrison III et. al. 2015), and exclusion netting. The latest method, designed by the French extension services in 2005 (Severac & Romet, 2007) to control the codling moth, has been tested also in Italy. In both countries, it has been proved that exclusion netting represent one of the most readily available tools for crop protection and an environmental friendly alternative to pesticides (Alaphilippe et.al 2016). As a matter of fact, this method is currently applied on several thousand hectares of orchards in both countries (Caruso et al. 2017) and other applications are currently being carried out in different parts of the world (Chouinard et al. 2016). The present study aimed at investigating exclusion nets as a potential strategy to prevent BMSB damage in pear orchards. The experimental activities, performed during 2016- 2018 included: 1) A laboratory trial was performed to evaluate the effectiveness of different net’s mesh size (mm 2.2x2.2, mm 4.0x2.5, mm 5.0x1.3, mm 7.0x3.0), against different instars (II, III, IV,V and adults). 2) field trials, conducted in commercial orchards in the province of Modena, using different types of net as follows: a. eight IPM orchards with “anti-hail nets”( mesh size of mm 7,0x3,0) in comparison with 8 uncovered orchards; b. eight IPM orchards with “whole orchard-nets” (made of an anti-hail net at the top with mesh size of mm 7,0x3,0, and the net whit mesh size of mm 4,5x2,0 on the perimeter), in comparison with 8 uncovered orchards; c. eight organic orchards with single-row nets (mesh size of mm 4,5x2,0) in comparison with 8 uncovered orchards. The results of laboratory trials show that (4.0x2.5) and (5.0x1.3) mm mesh size net, are able to exclude the most damaging instars, such as adults and older nymphs (IV-V instars). Nets with smaller mesh size (2.2x2.2 mm) exclude instar III also, with no significant efficiency gains. Therefore, the nets (4.0x2.5 and 5.0x1.3 mm), commonly used in exclusion netting and already widespread in the field, could be considered reliable for this type of application. The results of field trials indicated that:

PheroFip 2019 205

- “anti-hail net” alone reduced damage on pears about 50% compared to un-netted orchards but no differences were detected in the number of applied insecticide treatments (n. 6,5 treatments/year on average); - in “whole orchard-net” damage on pears was reduced about 80 % compared to un- netted orchards and insecticide treatments were reduced about 35%; - in “single-row net” fruit damage was reduced by almost 90 % compared to un-netted orchard in organic farms, where no effective insecticides are available/authorized. These results showed that exclusion netting systems are an efficient and sustainable tool in the framework of IMP and Organic programs for BMSB. However, being this species a very mobile insect, (Lee & Leskey, 2015), with five instars of different size able to move in narrow spaces, the efficacy of exclusion netting is not always close to 100%, and some additional insecticide treatments might be necessary. These observed positive results induced the Regional administrations of Northern Italian Regions to contribute financing the installation of exclusion netting with either whole or single-row nets in fruit orchard farms.

Key words: Brown marmorated stink bug, exclusion netting, integrated pest management, pear orchards, invasive insect

Acknowledgements This study was funded by the Emilia Romagna region within the Rural Development Plan 2014-2020 Op. 16.1.01- GO PEI-Agri - FA 4B, Pr. «Halys», coordinated by CRPV.

References Alaphilippe, A., Capowiez, Y., Severac, G., Simon, S., Sandreau, M., Caruso, S. & Vergnani, S. 2014: Codling moth exclusion netting: an overview of French and Italian experiences. Proceeding of IOBC meeting «Pome arthopods and Stone fruits» - Vienna (Austria) 6-9 October. Blaauw, B. R, Polk, D. & Nielsen, A. L. 2014: “IPM-CPR for Peaches: Incorporating Behaviorally-Based Methods to Manage Halyomorpha halys and Key Pests in Peach. Pest Management Science 71(11): 1513–22. doi:10.1002/ps.3955 Caruso, S., Vaccari, G., Vergnani, S., Raguzzoni, F. & Maistrello, L. 2017: Nuove opportunità di impiego di reti multifunzionali. L’Informatore Agrario n. 15, 57: 60. Chouinard, G., Firlej, A. & Cormier, D., 2016: Going beyond sprays and killing agents: exclusion, sterilization and disruption for insect pest control in pome and stone fruit orchards. Scientia Horticolture, 208 Maistrello, L., Vaccari, G., Caruso, S., Costi, E., Bortolini, S., Macavei, L., Foca, G., Ulrici, A., Bortolotti, P. P., Nannini, R., Casoli, L., Fornaciari, M., Mazzoli, G. L. & Dioli, P. 2017: Monitoring of the invasive Halyomorpha halys, a new key pest of fruit orchards in Northern Italy. Journal of Pest Science 88 (1): 37–47. DOI 10.1007/s10340-017-0896-2 Morrison, W. R., Lee, D. H., Short, B. D., Khrimian, A. & Leskey, T. C. 2016: Establishing the Behavioral Basis for an Attract-and-Kill Strategy to Manage the Invasive Halyomorpha halys in Apple Orchards. Journal of Pest Science 89 (1): 81–96. doi:10.1007/s10340-015-0679-6. Severac, G. & Romet, L. 2007: Alt’Carpo, contre la carpocapse travailler avec filets. Phytoma, 601: 10-14. Lee, D. H. & Leskey, T. C. 2015: Flight behavior of foraging and overwintering brown marmorated stink bug, Halyomorpha halys (Hemiptera: Pentatomidae). Bullettin of Entomological Research. Vol. 105, Issue 5, pp. 566-573.

PheroFip 2019 206

First approach to manage the invasive Halyomorpha halys in Italy: a three- year project

Maria Grazia Tommasini1*, Lara Maistrello2, Francesca Masino2, Andrea Antonelli2, Pier Paolo Bortolotti3, Roberta Nannini3, Stefano Caruso3, Giacomo Vaccari3, Luca Casoli3, Michele Preti4, Marco Montanari4, Matteo Landi4, Marco Simoni4, Stefano Vergnani5

1CRPV, Reasearch Centre Crop Production, Via dell’Arrigoni. 120, 47522 Cesena (FC), Italy 2Dip. Scienze della Vita, Università di Modena e Reggio Emilia, via G. Amendola 2, 42122 Reggio Emilia (RE), Italy; 3Consorzio Fitosanitario di Modena, Via Santi Venceslao 14, 41123 Modena (MO), Italy;4ASTRA Innovazione e Sviluppo Test Facility, Via Tebano 45, 48018 Faenza (RA), Italy 5OROGEL, Via Dismano 2600, 47522 Cesena (FC), Italy *[email protected]

Abstract Soon after the first detection in Italy of the invasive Halyomorpha halys (Stål), commonly named Brown Marmorated Stink Bug (BMSB), high losses were reported in Emilia Romagna region on fruit orchards, with up to 60% fruit damaged in integrated crop production and over 90% in organic management (Maistrello et al., 2017). Emilia Romagna is a very intensive fruit production region with over 56,164 hectares of fruit crops and 53,456 hectares of grapevine (ISTAT 2017) and BMSB created high concern in the whole agriculture sector. The fast spread and damage levels caused by BMSB forced farmers to increase broad- spectrum insecticide treatments, disrupting the Integrated Pest Management (IPM) strategies previously applied on pome fruits that included mating disruption and use of microbiological products (e.g. CpGV). To face this serious threat, farmers, technicians and researchers of public and private structures have started in 2016 a three-year project to identify new approaches to manage this pest. This project, called “HALYS”, encopasses a series of activities that included: insights on BMSB biology and monitoring techniques, agro- ecological studies, the biocontrol potential of native antagonists, evaluation on the use of physical barriers and on chemical products authorized in Italy. In particular, the project aimed to identify both prompt effective solutions as well as more environmental low impact solutions to control the BMSB infestations. Since 2016, a yearly based investigation on BMSB survival during overwintering and its biology, based on bugs kept outdoors, the dates and numbers of bugs successfully coming out of overwintering, the beginning of mating of both generations, the emergence of nymphs and adults were periodically communicated to Regional Plant protection Service and to integrated production advisors. These information together with data on egg to adult survival and development times are of immediate practical utility for the advisors and farmers, aiding in decision-making, and they will also be crucial for the elaboration of forecasting models. Based on the results achieved so far, none of the chemicals tested in the laboratory showed a satisfying efficacy to kill eggs whereas some effects were recorded on emerging nymphs. The semi-field trials showed a certain persistency with a residual effect at 3-7 days, of chemicals belonging to phosphorganics, neonicotinoids and pyrethroids, applied at 3 different timings against nymphs and adults of BMSB. In open field trials in pear orchards some chemicals show a certain mortality of BMSB in the short period, with nymphs more sensitive than adults to sprays. Besides, in agroecological studies an important influence of the surrounding agroecosystem was observed, together with the importance of overwintering

PheroFip 2019 207

and refuge places of BMSB (e.g., buildings, ornamental and wild trees, hedges) as source of infestation or re-infestation of fruit orchards. Innovative strategies based on ethology of the insect and optimizing/reducing chemicals application were also evaluated. In particular, the "IPM-CPR (Integrated Pest Management - Crop Perimeter Restructuring)” (Blaauw et al. 2014) was tested on pear orchards for two consecutive years. The strategy that focused on performing insecticide treatments only in the orchards edges, allowed a reduction of 40-50% of insecticidal inputs compared to standard applications on the whole orchard. Concerning the low impact solutions, exclusion netting either with antihail nets associated with border netting or single row nets, proved to be the most effective strategy to prevent BMSB infestation and damage on pear orchards compared to uncovered orchards (Caruso et al 2017). This technique allows a consistent reduction of chemical sprays and so far it appears to be the only one solution applicable in organic orchards to control the pest. Since BMSB is also present in the vineyards, but it is still not clear the damage that the pest can cause when grapes are destined to winemaking, the project also intended to verify the effects of the massive presence of these bugs during wine making, looking for variations on the quality of the red wine Lambrusco. Results achieved so far indicate no heavy variation on the chemical profile of the wine. Sensorial analysis showed the presence of some anomalous notes not belonging to the typical sensorial scent of Lambrusco, although it does not seem to be attributable to "bug smell". In general, the project HALYS gave a first crucial contribution to support farmers and IPM technicians of Emilia Romagna region to manage the BMSB infestations although many aspects remain to be investigated to find sustainable strategies for all the crops affected by this invasive pest.

Key words: Brown Marmorated Stink Bug, integrated control, pome fruits, grapevine

Acknowledgements This study was funded by the Emilia Romagna region within the Rural Development Plan 2014-2020 Op. 16.1.01- GO PEI-Agri - FA 4B, Pr. «Halys», coordinated by CRPV.

References Blaauw, B. R., Polk, D. & Nielsen, A. L., 2014: IMP-CPR for pheaches: incorporating behaviorally-based methods to manage Halyomorpha halys and key pests in peach. Pest Management Science 71(11): 1513-22.doi:10.1002/ps.3955 Caruso, S., Vaccari, G., Vergnani, S., Raguzzoni, F. & Maistrello, L. 2017: Nuove opportunità di impiego delle reti multifunzionali. Informatore Agrario 15/17 pag. 57-60 ISTAT http://agri.istat.it/jsp/dawinci.jsp?q=plC170000020000093200&an=2009&ig=1&ct=266 &id=15A|21A|30A (28 Sept 2018) Maistrello, L., Vaccari, G., Caruso, S., Costi, E., Bortolini, S., Macavei, L., Foca, G., Ulrici, A., Bortolotti, P.P., Nannini, R., Casoli, L., Fornaciari, M., Mazzoli, G.L. & Dioli, P., 2017. Monitoring of the invasive Halyomorpha halys, a new key pest of fruit orchards in Northern Italy. Journal of Pest Science 88(1): 37–47. DOI 10.1007/s10340-017-0896-2

PheroFip 2019 208

Habitat and tree species effects on Trissolcus japonicus (Hymenoptera: Scelionidae) detections in Virginia, USA

Quinn, N.F.1, Talamas, E.J.2, Leskey, T.C.3, Bergh, J.C.1

1Virginia Tech, Alson H. Smith, Jr. Agricultural Research and Extension Center, Winchester, VA, USA 2Florida Department of Agriculture and Consumer Services, Gainesville, FL, USA 3USDA Agricultural Research Service, Appalachian Fruit Research Station, Kearneysville, WV, USA

Abstract Native biocontrol agents in the USA have not adequately impacted populations of the invasive agricultural and nuisance pest, brown marmorated stink bug, Halyomorpha halys (Hemiptera: Pentatomidae). In Asia, the egg parasitoid, Trissolcus japonicus (Hymenoptera: Scelionidae), is a key natural enemy of H. halys. Multiple adventive populations of T. japonicus have been detected in the USA since 2014 and its geographic range appears to be expanding. Thus, to track the spread and changes in the relative abundance of T. japonicus over large geographic areas, the optimization of sampling methods and protocols are needed. Yellow sticky traps deployed in the mid-canopy of tree of heaven, a common host of H. halys, have yielded increasingly frequent T. japonicus detections in Frederick County, Virginia. Using this approach and mature female tree of heaven as the standard sampling unit, traps were deployed in hedgerows, at the forest edge, and in spatially isolated patches from May through September, 2018. To evaluate H. halys host tree effects, traps were also deployed from June through August in pairs of trees along forest edges in 2017 and 2018. Each pairing contained female tree of heaven and one of the following; hackberry, black locust, black walnut, or cherry. For both studies, T. japonicus captures were recorded weekly. Significantly more T. japonicus were captured in hedgerows than at the forest edge. Captures in isolated patches were numerically greater than at the forest edge, but not significantly different from the other two habitats. In 2017, T. japonicus was captured only in tree of heaven, although low captures resulted in no statistical differences between hosts. In 2018, it was captured in all host species but there were not significant differences between any host pair. In 2018, T. japonicus captures occurred from 11 May until 9 September, with distinct peaks in early July and early August. Results are discussed in relation to aspects of the foraging ecology of T. japonicus and implications for sampling.

PheroFip 2019 209

SESSION 7

INVASIVE PESTS: A CHALLENGE FOR IPM

B - SPOTTED WING DROSOPHILA AND OTHER INVASIVE

DIPTERANS

CHAIRPERSON: HOWARD THISTLEWOOD

CHAIRPERSON: HEIDRUN VOGT

PheroFip 2019 210

Seasonal changes in odour preferences of Spotted Wing Drosophila (SWD) and the impact of feeding and reproductive status

Tim Beliën1, Vincent Van Kerckvoorde1, Ammar Alhmedi1, Rik Clymans1, Eva Bangels1, Dany Bylemans1,2

1pcfruit, Zoology Department, Fruittuinweg 1, B-3800 Sint-Truiden, Belgium 2Department of Biosystems KULeuven, Decroylaan 42, B-3001 Heverlee, Belgium

Abstract The Asiatic vinegar fly Drosophila suzukii (Spotted Wing Drosophila, SWD) has recently invaded Europe, and immediately became a key pest of grapes, stone and soft fruits. Unlike other fruit flies that typically infest only overripe and rotten fruit, SWD females oviposit in ripening fruit, leading to considerable economic damage in a broad range of soft-skinned fruits. Here we present recent outcomes from applied research revealing important insights into SWD population dynamics and seasonal changes in odour preferences. Firstly, we demonstrated that the currently widely applied monitoring programs, with traps and liquid attractant based on apple cider vinegar (ACV), provide a distorted picture of the actual SWD population dynamics profile. Use of alternative attractants clearly showed that there is an underestimation of the SWD population development during the main soft and stone fruit production periods (late spring-summer). Secondly, we attempted to unravel the seasonal shifts in preference of the SWD population between fermentation cues (like ACV) and fruit odour. Differences in seasonal odour preferences between winter and summer morphs of SWD were observed in a two-year field experiment. Furthermore, our results indicate that feeding and reproductive status are stronger behavioural cues than the (winter) morphology itself, although present. This explains why winter and summer morphs preferred strawberry baited traps over ACV in periods of abundance. However when starved both morphs significantly preferred ACV, but this was intrinsic stronger for the winter morph than the summer morph.

Key words: Drosophila suzukii, monitoring, apple cider vinegar, fruit odour, winter morphology, choice assay

PheroFip 2019 211

Dissecting oviposition ‘decision making’ in Drosophila suzukii: a hierarchical sensory cascade dictates whether and how much to oviposit

Teun Dekker1, Paul Becher1, Emil Dekker1, Joelle Lemmen1

1Department of Plant Protection Biology, Swedish University of Agricultural Sciences, Sweden.

Abstract The invasive Drosophila suzukii has been much studied for its olfactory sensitivity to odors from its favored fruits. These odors are thought to be used in odor-mediated orientation of gravid females for oviposition hosts. However, how gravid females orient to potential hosts in relation to other sensory cues, has not been sufficiently spelled out. Here we demonstrate the overriding importance of visual, over olfactory cues, the latter largely acting as surround cues for environmental suitability. Data further support a stochastic model whereby oviposition rates are largely determined by residence time, which is modulated by proprioreception cues, modulated by olfactory cues with no interference of visual and intraspecific cues. The data support a different mode of odor cues in orientation, possibly with a more general significance, in which olfaction is more as contextual than orientational cue in defined behaviors.

PheroFip 2019 212

Improved ‘Attract & Kill’ control of Drosophila suzukii (spotted wing Drosophila SWD) with feeding enhancers

Van Kerckvoorde V1, Clymans R1, Bangels E.1, Alhmedi A.1, Thys T.1, De Ro M.3, Bonte J.3, Casteels H.3, De Clercq P.4, Bylemans D.1,2, Belien T.1

1pcfruit npo, Department of Zoology, Kerkom, Belgium, 2KULeuven, Department of Biosystems, Leuven, Belgium; 3ILVO, Plant, Department of Crop Protection - Entomology, Merelbeke, Belgium, 4Ghent University, Faculty of Bioscience Engineering, Department of Crop Protection, Laboratory of Agrozoology, Ghent, Belgium

Abstract The objective of this paper was to identify the most efficient feeding enhancer in the lab and confirm its ability to improve chemical control in an ‘Attract & Kill’ strategy of Drosophila suzukii, spotted wing Drosophila (SWD), in the field. Tested feeding enhancers were white sugar 0.2% (WS), brown sugar 0.2% (BS), Attracker® 0.2% (A), Combi-Protec® 0.2% (CP), Blossom Protect® 0.1% (BP) and a mixture of baker’s yeast 0.125% (Saccharomyces cerevisiae) and brown sugar 0.125% (BYBS). A wetting agent, Trend® 0.1% (T), was also included. The methods used in the lab were a capillary feeding assay, a petri dish assay (where the attraction of SWD to drops of feeding enhancer solutions was evaluated) and a fruit bioassay (where 10 SWD were exposed to fruit (untreated and dipped in cyanthraniliprole 0.05% with or without the feeding enhancers) in condiment cups and their mortality was scored after 16, 24, 48 and 72 hours). The field trial was performed in a sweet cherry orchard. It consisted of a random block design trial with four replicates of five trees that were sprayed weekly with water and spinosad 0.015% solo and combined with WS, CP and BYBS. The applications were made with a Stihl SR450 gasoline powered backpack sprayer at 1000 litres of water per ha leaf wall area. On regular intervals the number of larvae per 50 cherries was assessed by immersing the fruit in a 10% NaCl solution during 24 hours and sieving the larvae out prior to counting with a binocular. The results of the capillary feeding and petri dish assay revealed a shift in preference: at the beginning of these trials sugar-based feeding enhancers are preferred by SWD, but as the tests progress this preference shifts to protein based products. It is hypothesized that the SWD first ensure their survival by consuming a lot of sugar. Once this is need is fulfilled the focus shifts to anabolism through proteins. Both assays were however unable to identify the best feeding enhancer. In the fruit bioassay only BYBS significantly increased the mortality caused by cyanthraniliprole after 16 hours. BS and CP also increased the mortality after 16 hours, but this was not significant. After 24, 48 and 72 hours no feeding enhancer could improve the mortality in a significant manner. The wetting agent T significantly decreased the mortality at 16 hours and 24 hours. This drop in mortality was so pronounced that there was no significant difference any more with the untreated control. In the field trial only the mixture of backers yeast 0.125% and brown sugar 0.125% was able to improve the effect of spinosad on all three sampling dates but this was not significant. Statistical analysis was made with a Friedmann non-parametric and a Student-Newman-Keuls test (non-transformed variables, ∝=0, 05). In conclusion, the mixture of baker’s yeast 0.125% and brown sugar 0.125% was the best feeding enhancer, both in lab (significant) as in field (not significant). The wetting agent Trend 0.1% was revealed as a feeding ‘disturber’ in the lab (significant).

PheroFip 2019 213

SESSION 8

INTEGRATED PROTECTION OF FRUIT

CROPS

CHAIRPERSON: PETROS DAMOS

PheroFip 2019 214

First studies of Grapholita molesta insecticide resistance in field populations from Catalonia (NE of Spain)

Dolors Bosch1, Adriana Escudero1, Jesús Avilla2

1IRTA, Institut de Recerca I Tecnologia Agroalimentària, C/ Rovira Roure 191, 25198 Lleida, España; 2Universitat de Lleida, ETSEA, Departament de Protecció Vegetal i Ciència Forestal, C/ Rovira Roure 191, 25198 Lleida.

Abstract Grapholita molesta (Busck) (Lepidoptera: Tortricidae) is a classical key pest for nectarines and peaches in Spain. However, in Catalonia, since 2011 it has also become an important pest for apples, mainly in Girona (NE of Catalonia). At present, there is a progressive increase of the populations and the problem is extending to the entire productive area. G. molesta mating disruption is used to control the pest in Girona, where apples are the main crop, covering at this moment approximately 80 % of the 2675 ha. In Lleida, where there is a crop mixture, 9750 ha of apples, 10150 ha of pears and 19850 ha of peaches and nectarines, the mating disruption against G. molesta is widely used in peaches and nectarines, its primary hosts, but not in the rest of the crops. Due to the increase of populations the use of chemical control is usually necessaire requiring a very variable number of treatments, depending on the crop and the variety. Despite the treatments, control failures has been recorded during last years. Early detection of insecticide resistance is very valuable for a rapid implementation of a resistance management strategy to prevent the appearance or the extension of the problem. The susceptibility of four and one field populations collected in the monoculture area of peach and the mixed crop production area in Lleida, respectively, and one field population from Girona to eight insecticides were tested and compared with a susceptible laboratory strain (GM-S-Spain) mass reared at laboratory using a semi-artificial dehydrated diet, for more than 10 years. The diagnostic concentrations that produced approximately 90 % mortality in the susceptible population (LC90) were 0.7 for deltamethrin, 43.3 for phosmet, 2.8 for chlorpyrifos-methyl, 7.2 for thiacloprid, 6.2 for chlorantraniliprole, 9.3 for indoxacarb, 5.1 for methoxyfenozide and 1.6 for spinetoram. Methoxyfenozide and deltamethrin were the products with more field populations showing decreasing susceptibility, whereas none of the field populations had loss of susceptibility to the rest of insecticides. Just one of the field populations showed a resistant ratio higher than 10 to methoxyfenozide. More field populations should be tested but, according to these preliminary results, there is not a problem of insecticide resistance in the area, therefore is the best moment to implement a resistance management strategy.

Key words: Grapholita molesta, insecticide resistance, management, integrated control

PheroFip 2019 215

Low-residue fruit production in the Lake Constance region – Model orchard systems for integrated crop protection

Diana Zwahlen1*, Barbara Egger1, Esther Bravin1

1Agroscope, Competence Division Plants and Plant Products, Müller-Thurgau-Str. 29, 8820 Wädenswil, Switzerland *[email protected]

Abstract Fruit production in the Lake Constance region has to meet high standards. On one hand, consumers and retailers expect inexpensive fruits of high quality, while on the other hand they should be free of residues and produced in an environmentally friendly way. More and more frequent weather extremes and increasing regulatory restrictions on pesticide authorizations exacerbate this tension. Within the presented project, innovative and system-based strategies are investigated and further developed to determine how fruits can be successfully produced under these conditions. The project focuses on the combination of promising measures to reduce the use of pesticides in model orchards. Those measures include variety selection, rain cover systems, exclusion nettings, mechanical weed control and promotion of beneficial insects. The aim is to measurably reduce the use of pesticides in comparison to common standards (integrated pest management and organic production) and to produce fruits that show preferably no residues, while maintaining a high internal and external quality. Besides thorough investigations concerning the impact and possible side effects of each measure, special emphasis is also given to their cost-effectiveness by conducting cost-benefit analyses and profitability analyses. All investigated aspects are compared to common standards. In total, five model plants were built for the project in the Lake Constance area, two of them by Agroscope: one apple orchard in Wädenswil (Canton Zurich) and one cherry orchard in Wintersingen (Canton Baselland). Both orchards were finalized in spring 2018. In the apple orchard with the varieties ‘Gala Buckeye’ (standard) and ‘Bonita’ (scab- resistant), the contribution of rain covers to control diseases and of exclusion nettings to control winged insects, as well as the combination of those two measures, is investigated. These measures are complemented by mechanical methods for weed control and various elements for the promotion of beneficial insects. In addition to the potential of those measures to reduce the use of plant protection products, their impact on diseases, pests, beneficial insects, microclimate, tree growth, crop quantity and quality, shelf life and residues on the harvested fruits is explored in detail. The focus in the cherry orchard is to find an optimal netting strategy with regard to the period of netting. In Swiss cherry production, lateral exclusion nettings are already widely used in the control strategy of Rhagoletis cerasi and Drosophila suzukii. However, it remains unclear whether the management of pests and beneficial insects could be optimized by an improved netting strategy, which then could lead to a further reduction of insecticide use. For this purpose, the model orchard consists of four sections; for each section, the netting and rain cover can be opened and closed independently of each other. This allows the examination of up to four strategies per year and a direct comparison. In addition to the netting strategy, benefit and impact of a reduced fungicide strategy, mechanical weed control and different elements for the promotion of beneficial insects will be assessed.

PheroFip 2019 216

Through the holistic approach and the extensive investigations and evaluations of the five model plants, this project will make a decisive contribution to the further development of the Integrated Pest Management.

Key words: fruit production, low-residue, integrated pest management, model systems, rain cover, exclusion netting, mechanical weed control, beneficial insects

Acknowledgements The project is part of the INTERREG V program of the European Union, funded by the European Fund for Regional Development and the Swiss government. Further project partners are Kompetenzzentrum Obstbau-Bodensee (D), Hochschule Weihenstephan-Triesdorf (D) and Bildungs- und Beratungszentrum Arenenberg (CH), which evaluate one model orchard each, Landwirtschaftliches Zentrum St. Gallen (CH), which evaluates an IPM orchard that serves as a reference, as well as AGRIDEA (CH), Landwirtschaftliches Technologiezentrum Augustenberg (D) and Landwirtschaftskammer Vorarlberg (A).

PheroFip 2019 217

Towards improved integrated control of Anthonomus spp. weevils in pome fruit orchards

Eva Bangels1, Tim Beliën1, Rik Clymans1, Eva Bangels1, Tom Thys1, Dany Bylemans1,2

1pcfruit, Zoology Department, Fruittuinweg 1, B-3800 Sint-Truiden, Belgium 2Department of Biosystems KULeuven, Decroylaan 42, B-3001 Heverlee, Belgium

Abstract Since the withdrawal of broad spectrum insecticides like carbamates and organophosphates and the restricted use of neonicotinoids, weevils have become an increasing problem in pome fruit in Europe. In particular, the apple blossom weevil Anthonomus pomorum and the pear bud weevil Anthonomus pyri (syn. A. cinctus or A. piri), that used to be secondary pests, have developed into important pests for many apple and pear growers, respectively. Recently, also the pear blossom weevil Anthonomus spilotus was found to be present in several pear orchards in Belgium, causing considerable damage. Here we present an overview of recent monitoring data, field and laboratory trials, executed in the Belgian fruit growing area near Sint-Truiden. Based on the recent and historical monitoring data the phenology of these pest species in pome fruit orchards was characterized into detail. Laboratory trials on A. pyri indicated a high direct contact activity as well as a high indirect residual contact activity of the neonicotinoid insecticides thiacloprid and acetamiprid, while the anthranilic diamide insecticide cyantraniliprole reached only good control activity after exposure of the A. pyri weevils to spray residue on leaves. In a field trial, application of thiacloprid and cyantraliprole targeting the active A. pyri weevils in autumn also resulted in very high control efficacies, leaving opportunities for a successful combination of those insecticides with biological control by the parasitoid Scambus pomorum. This opens perspectives for control strategies in line with sustainable integrated pest management (IPM) principles.

Keywords: Anthonomus spp. weevils, insecticides, Integrated Pest Management, apple, pear

PheroFip 2019 218

Silver leaf in Norwegian Prunus domestica plum orchards

Jorunn Børve1, Venche Talgø1, Arne Stensvand1,2

1Norwegian Institute of Bioeconomy Research, P.O. Box 115, 1431 Ås, Norway, 2Norwegian University of Life Sciences, Universitetstunet 3, 1433 Ås, Norway

Abstract Incidence of silver leaf caused by Chondrostereum purpureum in European plum, is normally low in newly planted Norwegian orchards. However, two to four years after planting high incidences could be observed. Questions raised by the growers are how fast the development of silver leaf is and how likely it is that the infection happened already in the nursery. Inoculation experiments have been initiated in addition to observations of symptoms and signs in orchards, i.e. silver coloured leaves, dead twigs and trees, internal decay and formation of fruiting bodies. Rapid development of silver leaf was observed after inoculation on trees in a growth chamber. In a commercial orchard, 50% of the trees had silver leaf 2 years after planting. Similar incidences were also observed in older orchards. In orchards with more than one cultivar planted at the same time, cultivar differences were observed. Fruiting bodies could be observed on dead branches and trunks of trees with silver leaf symptoms, also before the trees were dead.

Key words: Chondrostereum purpureum, fruiting bodies, nursery infections

PheroFip 2019 219

PheroFip 2019 220